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International Journal of Food Properties

ISSN: 1094-2912 (Print) 1532-2386 (Online) Journal homepage:

Processing of Honey: A Review

R. Subramanian , H. Umesh Hebbar & N.K. Rastogi

To cite this article: R. Subramanian , H. Umesh Hebbar & N.K. Rastogi (2007) Processing
of Honey: A Review, International Journal of Food Properties, 10:1, 127-143, DOI:

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Published online: 31 Jan 2007.

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International Journal of Food Properties, 10: 127–143, 2007
Copyright © Taylor & Francis Group, LLC
ISSN: 1094-2912 print / 1532-2386 online
DOI: 10.1080/10942910600981708


R. Subramanian, H. Umesh Hebbar and N.K. Rastogi

Department of Food Engineering, Central Food Technological Research Institute,
Mysore, India

Thermal processing of honey eliminates the microorganisms responsible for spoilage.

Microwave heating, infrared heating, ultrasound processing, and membrane processing
have been explored as alternatives to conventional heat processing. Microwave heating pro-
vides a rapid method for achieving the desired level of yeast reduction with reduced thermal
damage. Infrared heating is not as rapid as microwave heating but desired results are
achieved in a relatively shorter duration (3 to 4 minutes) compared to the conventional
method. Membrane processing is an athermal process and very effective in the complete
removal of yeast cells from honey. Microfiltration and ultrafiltration could be employed to
produce enzyme-enriched honey besides clarified honey.

Keywords: Diastase activity, Honey, Hydroxymethylfurfural (HMF), Infrared heating,

Membrane processing, Microwave heating, Ultrasound treatment, Yeast count.

Honey, a natural biological product evolved from nectar and of great benefit to
human beings both as medicine and food, is consumed in every country of the world in
some form. Honey contains glucose, fructose, and water, in addition to small quantities of
proteins, minerals, organic acids, and vitamins.[1] It is liked for its characteristic flavor,
sweetness, and texture.
Honey extracted from combs and apiaries contains pollens, beeswax, and other
undesirable materials, besides yeast, that are to be removed for better product quality and
shelf life. Hence, honey is processed before packing in bottles or other containers. The
type of equipment used and steps followed in processing, however, depends upon the
scale of operation. Two important stages of honey processing are filtration and heating.
The separation of pollens, beeswax, and other materials are normally done through strain-
ing and pressure filtration. Heat or thermal processing of honey eliminates the microor-
ganisms responsible for spoilage and reduces the moisture content to a level that retards
the fermentation process. The general process flow diagram for conventional honey pro-
cessing is provided in Fig. 1. The physical separation of undesirable, suspended matter is
done before thermally processing honey.

Received 8 May 2006; accepted 30 August 2006.

Address correspondence to R. Subramanian, Department of Food Engineering, Central Food Technological
Research Institute, Mysore 570020, India. E-mail:






Fine Particles

Moisture reduction

Yeast count


Figure 1 Conventional method of processing honey.

The straining operation to remove suspended solids (including large wax particles)
is carried out either manually or by mechanical means. The method and the equipment
used for straining depend on the size of the operation. In small-scale operations, straining
is done using cloth or nylon bags, which are frequently cleaned to remove the suspended
particles. In large-scale operations, the straining operation is combined with the pre-
heating (up to 40°C) operation in a jacketed tank fitted with a stirrer.[2]

The strained honey is further processed using pressure filters. Typically a polypro-
pylene micro filter of 80 μm is used as a filter medium. The honey temperature is main-
tained between 50–55°C, which prevents the melting of the beeswax.[2] Large-scale
processors subject honey to coarse filtration, centrifugal clarification, fine filtration, and
blending, prior to filling.


The major problem faced by honey producers in tropical countries is its rapid deteri-
oration in quality due to fermentation. Honey generally contains osmophilic (sugar–tolerant)
yeast in greater or lesser amount and ferment, if the moisture content is high enough and

storage temperature is favorable. Lockhead[3] had reported that a raw honey sample con-
taining more than 20% moisture readily undergoes fermentation irrespective of the initial
yeast count. Moisture influences the rate of fermentation, granulation, and honey flavor.
Reduction of moisture content below 17% is considered to be a safe level for retarding
yeast activity. However, the chances of granulation increase with decrease in moisture
content of honey. The unprocessed honey tends to ferment within a few days of storage at
ambient temperature because of its high moisture content and yeast count. To prevent fer-
mentation, honey is heat processed before storage. Heat processing of honey eliminates
the microorganisms responsible for spoilage and reduces the moisture content to a level
that retards the fermentation process.
Honey has high viscosity (1.36 N·s/m2 at 25°C and 21.5% moisture)[4] that pose
problems in handling and processing. Viscosity of honey is influenced by several factors.
As honey is heated, it initially undergoes a very rapid decrease in viscosity up to 30°C,
beyond which the change in viscosity is much slower. Besides temperature and moisture
content, variations in viscosity are attributed to the composition of individual sugars and
to non-sugar and colloidal material. Mossel et al.[5] successfully described the sugar con-
centration dependence of the viscosity of honey samples with varying moisture contents
with a model that was originally developed to describe the viscosity data of various sugar
mixtures. Newton’s law of viscosity could adequately describe the flow behavior of honey
samples, and the temperature dependency of viscosity was found to follow the Arrehenius
model.[6] Yanniotis et al.[7] also reported the Newtonian behavior of honey after studying
the effect of moisture content on its viscosity at different temperatures. Generally, honey
is a Newtonian fluid but non-Newtonian behaviour has also been reported owing to the
presence of colloids (possibly proteins) and the polysaccharide dextran.[8] Recently,
Escobedo et al.[9] reported that honey samples at the 12th week of storage showed a strong
tendency to behaving as a non-Newtonian fluid owing to the presence of crystals that
changed the flow behavior. The authors rightly cautioned that such a change in flow
behavior in honey may be due to the nature of the test. Sopade et al.[10] studied the pump-
ing characteristics of Australian honey and proposed an equation for the prediction of
pressure loss in a pipeline of typical size at any temperature and flow rate. Bhandari et
al.[11] correlated the glass transition temperature (Tg) with critical viscosity and reported
that the critical viscosity is reached at a temperature 10 to 20°C above the Tg.
Crystallization is an undesirable property in handling, processing, and marketing,
except for certain purpose such as in the production of creamed honey. Glucose is the
principal component that crystallizes in honey as it exists in a supersaturated state.
Bhandari et al.[8] summarized some of the methods proposed to stop crystallization of
honey: storage at freezing temperature (−40°C), heat treatment to dissolve crystals and
crystal nuclei, removal of air bubbles, dust and pollen particles by filtration, filling at
higher temperatures (>45°C) to avoid air bubbles incorporation during filling, addition of
inhibitors such as isobutyric and sorbic acid, and adjusting the glucose to fructose ratios
or the water content. Ultrasound processing has also been reported for preventing crys-
tallization in honey, and the various attempts made are discussed under a separate head-
ing. Heating is a common method to control the crystallization. It helps to melt invisible
crystals present in honey. After melting all the crystals and nuclei, even the most crystal-
lisable honey can remain liquid for many months. Presence of air bubbles in the packag-
ing containers can provoke nucleation and crystallization of honey. Filling at higher
temperatures eliminates air bubbles and avoids air incorporation during packing due to
low viscosity.[8]

Although, honey is thermally processed to eliminate yeast, it could result in product

quality deterioration. Uncontrolled heating alters the parameters such as hydroxymethyl-
furfural (HMF) content and enzyme activity unfavorably. The initial HMF content in dif-
ferent honey types varies drastically and it depends upon the climatic condition of the
region besides other factors. Excessive amount of HMF has been considered evidence of
overheating, implying a darkening of color and a loss of freshness of honey. The starch–
digesting enzyme(s) of honey are also used as an indicator of honey quality because of
their sensitivity to heat. The major enzymes present in honey are invertase, amylase (dia-
stase), and glucose oxidase along with minute quantities of catalase and acid phos-
phatase.[1] The enzyme content in honey is measured as diastase activity and expressed in
terms of diastase number (DN). The European Regional Standard for honey[12] specifies a
minimum DN of 8 in processed honey.


The conventional process involves preheating to 40°C, straining, filtering /clarification,
and indirect heating of filtered honey at 60–65°C for 25–30 minutes in a tubular heat
exchanger followed by rapid cooling in order to protect its natural color, flavor, enzyme
content, and other biological substances.[2] Studies have shown that heating honey at 63,
65, and 68°C for 35, 25, and 7.5 minutes, respectively can destroy the yeast cells
completely.[13] There is a lack of literature about the application of high-temperature
short-time (HTST) heating treatment on honey. Tosi et al.[14] reported that a mild HTST
treatment condition, typically heating at 80°C for 60 seconds in the transient stage and
30 seconds in the isothermal stage, destroyed all microorganisms responsible for quality dam-
age without spoiling honey. Temperature beyond 80°C through 140°C, at very short times,
did not seem to cause deleterious effects on honey as measured by HMF and diastase activity.
White et al.[15] reported the effect of storage and processing temperatures on honey
quality. Samples with high, intermediate and low diastase activity were stored at different
temperatures (20–70°C). The half-life time of diastase and invertase activity, estimated
using the empirical equations indicated the direct relationship with temperature. Singh and
Bath[16] studied the relationship between heating and HMF formation in different types of
honey. Heating temperature and time showed significant effect on HMF formation. They
observed a large difference in HMF formation between different types of honey and
used second order polynomials to effectively predict the HMF formation in different types
of honey. Effects of thermal treatment on HMF content in honey was also studied by Tosi
et al.[17] and reported that the kinetics of HMF formation did not depend on the initial
HMF concentration in honey. They also reported that during thermal processing, the time-
temperature combination is very crucial for maintaining the HMF level below the maxi-
mum permissible limit. Gupta et al.[18] studied the influence of different treatments and
storage conditions on some physicochemical characteristics and sensory qualities of
Indian honey. The color of the honey was significantly affected by the storage temperature
and period with maximum deterioration at a storage temperature of 40°C. However, gran-
ulation was completely eliminated in honey stored at 40°C due to melting. Evaluation of
honey samples stored for six months showed comparatively higher overall sensory score
for unheated honey stored at 5°C (Table 1). Visquert et al.[19] investigated the effect of
thermal processing on honey quality during storage. Honey was processed at 35 to 85°C
for 1–672 hours, depending on the chosen processing temperature and evaluated with
respect to HMF content, acidity, electrical conductivity, and moisture content. Thermal

Table 1 The effect of different treatment on the sensory characteristics of honey after six months of storage.[18]

Treatment Color Consistency Taste Aroma qualities

Room temperature (7–30°C)

Unheated 18.8 16.7 17.3 17.8 17.8
Unheated (KMS) 17.3 16.5 17.6 17.1 18.2
Heated 15.8 17.2 16.1 16.7 15.6
Temperature 5°C
Unheated 19.3 15.4 19.2 18.9 19.6
Unheated (KMS) 18.6 15.0 18.6 17.5 18.4
Heated 16.0 16.2 16.5 17.6 15.5
Temperature 40°C
Unheated 14.4 19.1 13.8 14.0 1.37
Unheated (KMS) 14.0 18.7 14.2 13.5 1.39
Heated 14.2 19.0 13.9 14.2 1.38

processing increased the HMF content of honey considerably. However, acidity, electrical
conductivity, and moisture content were unaffected by thermal processing and during sub-
sequent storage.
Central Bee Research and Training Institute, Pune had developed a honey process-
ing plant to handle Indian varieties of honey, which have very high moisture content.[20]
The plant having the capacity to process ∼40 kg/h, could reduce the moisture content by
∼8–10%. The constructional features of the plant include cartridge type micro filters, pro-
cessing tanks with helical coil heat exchangers, and falling-film evaporator.
In recent times the demand for better quality honey is on the rise as honey is being
consumed now for its health benefits. So, efforts are being made to look for alternatives to
conventional thermal process, which can produce better quality honey. Application of
microwave, ultrasound, and infrared heating of honey has been reported and claims have
been made on the improved product quality. Microwave and infrared heating have gained
popularity in food processing over conventional heating owing to their inherent advantage
of rapidity and better-quality product. Pressure-driven membrane processes are remark-
ably simple allowing ambient temperature operation and requiring less energy. Membrane
technology has the potential in replacing or complementing some of the traditional meth-
ods of processing, as well as in developing new products.


The application of microwave heating is well known in the food industry, particu-
larly for tempering, blanching, drying, and pasteurization of food material. Microwave
heating is greatly affected by the presence of water in foods, as water is the major absorber
of microwave energy in food, consequently, the higher the moisture content, the better the
resultant heating. In contrast to conventional heating, microwaves penetrate the material,
interact with it, and generate heat leading to its rapid heating. Materials containing polar
molecules, such as water, are rapidly heated when exposed to microwave radiation due to
molecular friction generated by dipolar rotation in the presence of an alternating electric
field. It is also reported that dissolved sugars are the main microwave susceptors in high
carbohydrate foods and syrups.[21] Since honey contains a substantial amount of water

(18–24%), as well as large amounts of dissolved sugars (70–80%), microwave radiation

could be effectively used for heating honey.
The processed honey containing yeast cells could be safely stored at room tempera-
ture provided the count is apparently insufficient to initiate fermentation. Ghazali et al.[22]
studied the effect of microwave processing of starfruit honey for its storage stability. Their
study showed that the fairly short time taken to reach the required processing temperature
ensured little change in chemical properties. Honey was heated to 71°C using a micro-
wave oven and stored at two different storage conditions, room temperature (28 ± 2°C)
and 4°C for 16 weeks. The physicochemical properties of unheated and heated honey were
measured before and during storage (Table 2). Spoilage was noticed in unheated honey
(control), irrespective of the storage temperature. Heated samples were more resistant to
spoilage. The spoilage of honey was attributed to the yeast count in honey, which was
much higher (1.02 × 105 cfu/g) in the unheated honey compared to heated honey sample
(5.90 × 102 cfu/g). No appreciable variation in yeast count was noticed during storage of
heated honey. Granulation was not observed in honey samples that were heated before
storing. Storing of unheated honey at room temperature was also free from granulation in
agreement with the general observation related to dextrose-to-water ratio and storage tem-
perature. Darkening of honey color was observed during storage in both heated and
unheated honey from light golden color to golden brown. However, the honey stored at
4°C was considerably lighter in color than honey stored at room temperature. Heated
honey was darker than the unheated honey, whatever the storage temperature. Heating did
not alter the moisture content of honey (20.8%). Also, no noticeable change in ash, nitro-
gen contents, pH, and acidity was observed in microwave heated honey sample. Heat pro-
cessing of honey led to a 37.5% loss in diastase activity. Storage at 4°C had no effect on
subsequent activity. However, room temperature storage led to further loss (33%) in activ-
ity. Heating did not show any effect on the sugar contents. However, the concentration of
glucose, maltose, and sucrose changed during storage depending on the storage tempera-
ture and whether the sample had been heated or not.
Bath and Singh[23] studied the effect of microwave heating on HMF formation and
browning in two types of honey (Helianthus annuus and Eucalyptus lanceolatus). The for-
mation of HMF and browning increased with microwave power levels as well as with
heating duration, the former showing greater effect. Both the types of honey differed sig-
nificantly with respect to HMF formation and browning under similar microwave heating
conditions, which was attributed to the difference in chemical composition of honey.

Table 2 The physicochemical properties of unheated and microwave heated starfruit honey before and after
storage for 16 weeks.[22]

Storage temperature

Property Unheated Heated 4°C Room temperature

Moisture (%) 20.80 20.80 21.00 (22.00) 20.80 (21.20)

Diastase number 4.00 2.51 2.32 (3.28) 1.69 (2.58)
Fructose (%) 32.10 32.12 32.10 (32.07) 30.08 (30.12)
Glucose (%) 30.14 30.10 30.02 (24.75) 26.85 (24.90)
Maltose (%) 2.31 2.30 2.42 (5.33) 5.47 (7.47)
Sucrose (%) 5.96 6.01 5.92 (5.48) 1.67 (0.00)
Yeast (cfu/g) 1.02 × 105 5.90 × 102 4.00 × 102 3.45 × 102

Values indicated in the parentheses are for unheated honey.


Hebbar et al.[24] conducted studies on microwave heating in a micro-convective

oven (2450 MHz, maximum power of 800 W). Experiments were carried out at different
power levels (PL) ranging from 10 to 100 (175–800 W) and for different heating periods
from 15 to 90 seconds. The extent of change in properties (HMF, diastase activity, mois-
ture content, and yeast count) mainly depended on the power level (power intensity) and
duration of heating. The changes in these properties were prominent in samples that were
heated at higher power levels and for longer durations. The peak temperature attained by
the sample depended on the power level used as well as duration of heating.
The reduction in yeast count was observed to the extent of commercially acceptable
level (< 500 cfu/mL) at power levels of 10, 30, and 50 when the samples were heated for
more than 45 seconds. At higher power levels of 70 and 100, heating duration of 30 and 15
seconds, respectively, was sufficient to achieve the same level of reduction in yeast count.
A semi-log plot of yeast count reduction ratio (ratio of yeast count at any given time to ini-
tial count) with time at different power levels is shown in Fig. 2a. The reduction in yeast
count was rapid, generally during the first 20 to 30 seconds, and the rate of yeast reduction
was directly related to the input power intensity. The reduction of yeast count is attributed
to the rapid increase in sample temperature due to microwave exposure, leading to the
rupture of yeast cell walls.
The increase in HMF value was gradual with heating duration at power levels of 10,
30, 50, and 70. But HMF level increased sharply in the samples heated for a longer dura-
tion at the maximum power level of 100. However, these values were far below the maxi-
mum permissible statutory level of 40 mg/kg of honey.[4] Variation of HMF with duration
of heating at different power levels is shown in Fig. 2b. The trends in the variation of
HMF values of the samples clearly depicted the sensitivity of honey to the period of heating
and temperature (power level).
Heating affects the enzyme activity and the diastase activity showed a decline with
heating under all conditions employed. The reduction in diastase activity with heating
duration at different power levels is depicted in Fig. 3a. Long heating periods of 60 to 90
seconds duration at power levels of 30, 50, and 70 reduced the diastase activity of honey
by ∼50% of its original value. At a power level of 100, heating above 45 seconds resulted in
reduction of the diastase activity to a level lower than the minimum permissible DN of 8.[12]
Heating also led to a reduction in moisture content above 9% at power levels of 50,
70, and 100 when the samples were heated for 60 seconds. Fig. 3b indicates the reduction
in moisture content with heating duration at different power levels. Larger reduction in
moisture content was not observed at lower power levels. The final moisture content in
most of the samples was in the range of 19.8 to 21.2%, which is below the acceptable level
(22%) for commercial processed honey.
Though different combinations of time and microwave power level could be used to
achieve the commercially acceptable level of yeast reduction in honey, it is equally impor-
tant to take the peak temperature attained by the sample into consideration. Heating honey
above 90°C results in caramelization of sugar.[25] Also it seems that this has a direct bear-
ing on the increase in HMF value and loss in diastase activity. Therefore, it is beneficial to
achieve the desired yeast reduction by choosing any suitable combination of power level
and duration that will maintain the temperature of honey well below 85–90°C.
Among the various selected combinations, higher power level and shorter duration
seems to be better than lower power level and longer duration. At power level 100
(power intensity 16 W/g), heating for 15 seconds resulted in substantial reduction in
yeast count (450 cfu/mL), lower HMF value (3.8 mg/kg) and higher retention of

Time (s)

0 20 40 60 80 100

P L10
-1 P L30
P L50
P L70
ln( A /A 0 )

P L100






PL 10
In c re a s e in H MF (%)

PL 30
PL 50
200 PL 70
PL 100




0 20 40 60 80 100
Time (s)


Figure 2 (a) Reduction in yeast count and (b) Increase in HMF with time at different power levels (PL) of
microwave heating.[24]

PL 10
PL 30
PL 50
PL 70
R educ tion in dias tas e

PL 100
ac tivity (%)





0 20 40 60 80 100
Tim e (s)



R educ tion in mois ture c ontent (%)

PL 10
PL 30
PL 70
PL 100

0 20 40 60 80 100
Time (s)

Figure 3 (a) Reduction in diastase activity and (b) Reduction in moisture content with time at different power
levels (PL) of microwave heating.[24]

diastase activity (DN 12). Also, the desired reduction in yeast count was achieved with
least undesired changes in the sample at a much lower processing temperature (54°C).


Infrared heating of food is gaining popularity due to its transient response, significant
energy savings over other thermal processes and ease of construction of hybrid systems
with convective and conductive heating sources.[26] Infrared heaters provide high rates of
energy input to the material surface and the radiant heat flux penetrates the material to a
depth, which depends on the nature of the material and the wavelength of the incident
radiation.[27] Sugar and water are the two major constituents of honey and both have good
absorption bands in the thermal radiation region.[26] The above factors could be successfully
utilized for efficient processing of honey.
Hebbar et al.[24] conducted studies on infrared heating in a near infrared (NIR) batch
oven (locally fabricated) fitted with lamps (1.0 kW, peak wavelength 1.1–1.2 μm). In
these experiments, the samples were heated continuously for 2, 3, 4, 5, and 8 minutes.
HMF, diastase activity, moisture content and yeast count in these samples were analysed
(Table 3). In all the cases, heating caused substantial reduction in yeast count. Heating for
5 minutes resulted in a product temperature of 85°C, HMF increase of 220%, and 37%
drop in enzyme activity. When the samples were heated for 8 minutes, no viable colony
forming units of yeast were noticed. However, the diastase activity drastically reduced in
these samples, clearly indicating excessive heating of honey that also showed up in a very
high product temperature (110°C). A heating period of 3 to 4 minutes was adequate to
obtain a commercially acceptable product, which met all the statutory requirements of
quality in terms of HMF (≤40 mg/kg), diastase activity (DN ≥ 8), moisture content, and
yeast count.


Granulation or crystallization is a natural tendency of honey and this common
phenomenon is a serious problem that affects marketing of honey. Crystallization of
honey is a complex process controlled by a number of factors acting simultaneously
and sometimes in a contradictory fashion. Concentration and super-saturation of the
major constituents (levulose, dextrose, and sucrose) and the minor ones (proteins and
dextrins), the presence of colloidal particles (nuclei), and temperature with its varying and
contradictory effects are some of the more important factors involved.[28] Traditionally,

Table 3 Continuous heating of honey with infrared radiation.[24]

Time Temperature Moisture Yeast count HMF Enzyme

S. No. (min) (°C) (%) (cfu/mL) (mg/kg) activity (DN)

Control 21.8 7000 2.0 16.6

1 2 47 20.2 500 3.2 13.8
2 3 61 19.8 300 3.6 12.4
3 4 74 19.8 200 4.6 11.6
4 5 85 19.2 150 6.5 10.5
5 8 110 18.2 Nil 7.9 Traces

honey is heated to temperatures of 77°C to kill yeasts and to delay crystallization.[29]

Heating affects the delicate flavors of honey, therefore alternate methods by means
other than heating such as ultrasonic, freezing, and chemical inhibitor treatments were
attempted for preventing crystallization in honey.[28] Ultrasonic waves are sound waves
with a higher frequency than the maximum that can be sensed by the human ear.
These waves when transmitted through liquid medium cause mechanical and thermal
changes in the material through which they pass, and also induce changes in unicellular
Kaloyereas[31] reported first in 1955 that high frequency sound waves (9 kHz)
eliminated the existing crystals and retarded further crystallization in honey. Ultrasound
processing destroyed most of the yeast cells that were present in the honey, and those that
survived had lost their ability to grow. No crystals were observed in ultrasound treated
honey and inhibited granulation for a period (15 months at 16°C) comparable to heating
the honey.[28] One disadvantage of this method was that exposure times of 15 to 30 min-
utes were required with cost implications.
Liebl[29] proposed an improved method for preventing the granulation by exposing
the honey to ultrasound waves of a much higher frequency (18 kHz) that drastically
reduced the liquefaction time to less than 30 seconds. This patented process was designed
to work at lower processing temperature (33°C) facilitating greater retention of aroma and
flavor along with huge savings on cost of energy compared to the conventional processing
involving heating and cooling steps. Studies were carried out at a considerably higher scale
(liquefaction of ∼1500 kg of honey/h) to demonstrate the claims on the cost effectiveness
of the process.
Thrasyvoulou et al.[30] studied the effects of ultrasonic waves on the quality of
honey focusing on some of the chemical characteristics. Crystallized honey samples
(100 g each) were liquefied by ultrasonic waves at 23 kHz and compared with conven-
tionally heated (water bath heating; 60°C for 30 minutes) and untreated samples. The
complete liquefaction of honey (5 samples each of blossom honey and honeydew
honey) required 18 to 25 minutes of ultrasound processing. Accordingly the energy
required for liquefaction varied from 0.1056–0.1466 kWh, and the maximum temper-
ature attained by the samples from 76–82°C. The variation in the time required for liq-
uefaction was attributed to the original granulated condition and the nature of
The combined effect of temperature and processing time resulted in increase in
HMF level. The average increase in HMF content was significantly low (86%) in samples
liquefied by sonication compared to samples liquefied by heating (129%). Ultrasonic
energy negatively affected the distaste activity of samples. The average decrease of dia-
stase activity was 16% after ultrasonic treatment and 23% after heat treatment. The influ-
ence of factors other than sonication or heat and typical behavior of individual samples
could also affect diastase activity. Moisture content, electrical conductivity, and pH were
not significantly affected by the treatments. The ultrasonic and heat treated samples were
stored at 25 ± 4°C and there was no significant difference in their recrystallization time.
The ultrasound treated samples remained in the liquefied state for 344 ± 39 days and heat
treated samples for 282 ± 86 days.[30]
Liquefaction by ultrasonic treatment affect the honey quality to a lesser extent com-
pared to heat treatment. It may be desirable to undertake studies on the effect of ultrasonic
treatment on a wide range of frequencies and also by eliminating the associated thermal
effect due to temperature rise.


Membrane processing is an athermal process and an alternate approach to the
conventional process. It is difficult to completely destroy the microorganisms present in
honey by traditional thermal processing methods practiced by the industries. Besides,
thermal processing results in reduction in enzymatic activity. Anticipated benefits of
membrane processing of honey are no cloudiness or sedimentation/granulation in
the product, reduced viscosity, commercially sterile product and consistent quality
The applications of ultrafiltered honey in gel formulations, cosmetics and pharma-
ceutical preparations besides its use as sweetener in tea/coffee and fruit beverages have
been reviewed by National Honey Board.[32] Itoh et al.[33] observed that poor rising of
sponge cakes (castilla) and sediment formation in fruit juices (lemon, grape, and apple),
when honey is used as an ingredient, was due to the water-soluble proteins (enzymes)
present in the honey. Further, they showed that ultrafiltration (UF) of honey completely
eliminated these problems. Besides, UF membrane with a molecular weight cutoff
(MWCO) of 10 000 could completely remove the microorganisms present in honey,
rendering it as a microbiologically safe ingredient.
But the ultrafiltered honey is devoid of desirable enzymes and proteins, and hence,
cannot be regarded for applications related to health foods. The major enzymes present in
honey are amylase or diastase (α-amylase), invertase (α-glucosidase), and glucose
oxidase. Diastase and invertase are nutritionally important enzymes present in honey. Dia-
stase hydrolyses carbohydrates for easy digestibility while invertase hydrolyses sucrose
and maltose.[1] Glucose oxidase is another important enzyme in honey that catalyses
glucose to form gluconic acid and hydrogen peroxide.[1] Generally, honey is well known
for antimicrobial activity against a number of microorganisms, probably due to the
presence of high levels of tetracyclines, phenolic compounds, and hydrogen peroxides.[34]
Sato and Miyata[35] attributed the antimicrobial property of honey predominantly due to
hydrogen peroxide, which allows its use in the treatment of wounds and gastrointestinal
diseases such as dyspepsia, bacterial gastroenteritis, gastric, and duodenal ulcers.
White and Kushnir[36] reported the approximate molecular weights for major honey
enzymes as 24 000 Da for amylase and 51 000 Da for α-glucosidase. Bergner and
Diemair[37] reported that glucose oxidase exhibits the highest molecular weight (>100 000 Da)
among the enzymes present in honey. The enzyme content in honey is measured as dia-
stase activity, which includes only amylases and not other enzymes such as invertase and
glucose oxidase. It is also preferable to measure the amylase content owing to its lower
molecular weight for the estimation of enzyme retention/rejection by the membranes.
The honey processors practicing membrane technology do not divulge technical
information as they are regarded as trade secretes directly linked with their commercial
success. There are very few reports available on membrane processing of honey in the lit-
erature. Itoh et al.[38] used 7000, 30 000, and 80 000 MWCO membranes in honey pro-
cessing and reported that bacteria and protein could be eliminated in honey using UF
membranes. Itoh et al.[39] also assessed the performance of UF process for honey with var-
ious processing parameters in a cross-flow membrane apparatus. The authors showed that
the total permeation flux, as well as permeation rate of sugar increased with increase in
operating temperature and increased dilution of honey at constant feed flow velocity and
applied pressure. Permeate flux also increased with increase in MWCO of the membrane
(10 000, 30 000, and 150 000). The authors also measured the water activity at various

sugar concentrations in honey for 4 different types of honey, which suggested that the
sugar concentration shall be greater than 35.5% to maintain the water activity below 0.94
so as to inhibit multiplication of microorganisms. Protein content in the permeate
decreased with decrease in MWCO of the membrane (500 000, 150 000, and 30 000) and
the permeate of 30 000 MWCO membrane did not contain any protein.
Barhate et al.[40] examined the rejection of enzymes in honey (50% diluted with
water) with various MWCO UF membranes and the effectiveness of these membranes in
eliminating yeast cells. Besides, attempts were made using membrane technology to pro-
duce a honey that is free of microorganisms and suspended matter, but containing a signif-
icant concentration of enzymes.
UF membranes (20 000, 25 000, 50 000, and 100 000 MWCO) completely removed
yeasts (Table 4). The sugar content in the feed was 220 mg/mL and did not change during
the UF process. The conventional heat treatment process used for honey is effective for
the inactivation of yeast but not for the other heat resistant organisms. It is reported that
microfiltration (MF) membrane with a pore size of 200 nm would remove the viable
microorganisms completely and as such could be used for sterilization.[41] Therefore, UF
membranes would be useful in eliminating other microorganisms besides effectively
removing the yeast cells present in the honey.
There was no diastase activity found in the permeates of UF membranes (Table 4).
The rejection of lower molecular weight enzymes gave a clear indication that enzymes of
higher molecular weights were also rejected by these membranes. These results imply that
there are reasons other than pore size of the membrane for the rejection. A secondary layer
formed on the upstream side of the membranes seems to be responsible for the complete
rejection of enzymes. Secondary layer or in other words dynamic membrane formation is
a process in which an active layer is formed on the membrane surface due to adsorption
and other deposition phenomenon of the substances contained in the feed being processed.
Such layers may also influence the rejection of other solutes present in the feed and can be
used advantageously in the process.[42]

Table 4 Rejection and permeate flux of different MF/UF membranes.[40]

Membrane Enzyme Rejection Av. flux

pore size/MWCO Sample activity (DN) Yeast (cfu/mL) of enzymes (%) (kg/m2·h)

MF membranes
25 nm Feed 5.3
Permeate 1.9 ND 66 ND
100 nm Feed 5.5
Permeate 2.6 ND 54 1.49
450 nm Feed 4.8
Permeate 3.6 ND 26 1.77
UF membranes
20 000 Feed 4.9 375
Permeate 0.0 0 100 ND
25 000 Feed 4.9 500
Permeate 0.0 0 100 0.90
50 000 Feed 5.0 600
Permeate 0.0 0 100 1.00
100 000 Feed 5.5 325
Permeate 0.0 0 100 1.15

ND, Not Determined.


Permeate flux is an important factor that determine the economics of the membrane
processes. In a situation like this, permeate flux is also influenced by the dynamic active
layer besides the flow resistance offered by the porous membrane, which is primarily con-
trolled by the pore size. The dynamic layer offers benefits for the overall processing of
honey since the permeate flux can be improved significantly by adopting higher MWCO
membranes without making any compromise in the rejection requirements of the process.
As expected MF membranes gave greater permeate flux and lower rejection com-
pared to UF membranes (Table 4). The rejection of enzymes was between 26 to 66%
depending on the pore size of the membrane. The rejection decreased as the pore size

Enzyme Enriched Honey

The enzymes present in honey, is the special reason for its wide spread use in nutri-
tion, therapeutic, and health related applications.[35] During the UF process, these enzymes
are retained in the retentate fraction limiting the use of ultrafiltered honey (with zero dia-
stase activity) in the above applications. A different processing strategy for the production
of enzyme enriched honey is possible using a combination of MF and UF membranes,
which is outlined in Fig. 4. The use of MF membrane (pore size 100 nm) in the process
will ensure elimination of yeast as well as other microorganisms while retaining the nutri-
tional factors in the product stream (permeate) at a higher permeation rate through the sys-
tem. An appropriate UF membrane (20 000 MWCO) will fractionate the microfiltered
honey into two fractions namely enzyme enriched honey and regular ultrafiltered honey.
By adopting this method, it is possible to enrich enzymes in honey to the extent of 2.2
fold. Higher enrichment of enzymes can be achieved by altering the volume concentration
ratio (VCR) in the membrane process. Such an enriched product with enzymes may find
use in very special applications related to health. These studies were conducted in batch
mode with stirred membrane cells, and the performance data obtained could form a basis
for conducting pilot scale process assessment before commercialization.

Concentration of Membrane Processed Honey

Honey is very viscous and is required to dilute with water before membrane pro-
cessing. In the case of UF and MF, there is no change in the water content during the pro-
cess. Though water removal is an expensive process step, the added water from the
membrane-processed honey is to be removed to obtain the original consistency. Vacuum
concentration at low temperatures is desirable to minimize the thermal damage to the
product. Despite the loss in the evaporation step (Table 5), there is an overall enrichment
of enzymes in the UF retentate fraction in the total process (Fig. 4).

Microwave heating can be effectively used for thermal processing of honey, as it
provides a rapid heating to achieve the desired results for long-term storage. Infrared heat-
ing is not as rapid as microwave heating but the desired results are obtained in a relatively
shorter period of 3 to 4 minutes offering advantages over the conventional method. Fur-
ther studies in the area of microwave and infrared heating of honey are needed to establish
the relationship between various processing conditions and honey quality in continuous

(Diluted honey at 1:1)

Pore size - 450 nm

Retentate (DN=3.6)
Pore size - 100 nm

Retentate (DN=3.6)
MWCO - 20000

Retentate Permeate
(DN=14.4) (DN=0)

Vacuum Vacuum
Concentration Concentration

Filtered enzyme Ultrafiltered honey

enriched honey (DN=0)

Figure 4 Schematic diagram of membrane processing of honey.[40]

Table 5 Quality variations during concentration of membrane processed honey.[40]

Operating conditions

Batch Temperature Absolute Loss in diastase Increase in HMF

size (g) (°C) pressure (mbar) activity (%) content (%)

250 50 50 20.8 5.2

400 57 98 27.0 9.0

flow systems to reach the industrial application level. Ultrasound processing destroys
most of the yeast cells present in the honey, besides eliminating the existing crystals and
retarding further crystallization in honey. Although this approach was first reported in
1955, projected benefits have not been realized even after 5 decades.
The UF membranes completely reject enzymes and totally eliminate yeast cells in
honey. Although scientific data is scanty, UF membranes are in commercial use to pro-
duce clarified honey. A combination of MF and UF membranes in the process provides a
scope to produce enzyme-enriched honey besides the regular clarified honey.

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