Vol. 51 No.

1 2007

Journal of Apicultural Science

23

HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

(HPLC) STUDY OF SUGAR COMPOSITION IN SOME
KINDS OF NATURAL HONEY AND WINTER STORES
PROCESSED BY BEES FROM STARCH SYRUP
Helena

Rybak-Chmielewska

Research Institute of Pomology and Floriculture, Apiculture Division, Department of Bee Products,
24-100 Puawy, Kazimierska 2, Poland. E-mail: helena.chmielewska@man.pulawy.pl
Received 02 November 2006; accepted 14 February 2007

S u m m a r y
Sugar composition was determined in three groups of products: starch (maltose) syrup produced
in Poland from wheat starch (for several years beekeepers have been trying to use it as bee food),
winter store made from the syrup by bees and honey. Winter stores were analyzed in three subgroups: subgroup 1 material collected from honeycombs of bee colonies two months after the
syrup was fed to the bees in autumn (end of October of 2004); subgroup 2 material collected from
the same bee colonies following seven months of feeding (spring of 2005); subgroup 3 samples of
winter stores which crystallized in the combs collected most of the time in early spring of 2006
(sent in by beekeepers from different regions of Poland from the season of 2005/2006).
Sugar content assays were made by HPLC with a refractometer detector according to
Bogdanov et al. (1997).
The following sugars were assayed and compared: glucose, fructose, sucrose, maltose, isomaltose, turanose and trehalose.
Significant differences were found for the contents of individual sugars in the stores made from
syrup vs. those in the honey. Routine HPLC assays of sugars can be helpful in the identification of
products made by bees from maltose syrups. The main distinguisher for those products was fructose
content (lower than that in honey by 32%), high maltose content (over 5%) and low fructose to glucose (F/G) ratio (0.76 when averaged across subgroups) whereas in honey samples it was 1.18 on
average. The lowest values of that parameter were found for rape honey averaging 0.98%.
Another problem explained by the study was that of the crystallization of stores processed from
maltose syrup following their depositing in the combs as winter storage. The crystallization occurred only in part of the apiaries that were fed maltose syrup in the season of 2005/2006. It was
found in that group of samples that fructose content was significantly lower and that of glucose significantly higher than in the remaining ones (non-crystallized or partly crystallized). According to
Ohe von der and Schnberger (2000) the critical point for at which a solution becomes saturated with glucose is 32 g/100 g and once that concentration is exceeded crystallization occurs. In
the examined samples the concentration of glucose averaged as much as 38.98%. It caused the sugar
to crystallize already in the comb cells. In the apiaries in which crystallization of maltose syrup derived stores occurred there were conditions which favoured enzymatic hydrolysis of complex sugars
to simple ones. Of particular importance here is the breakdown of maltose which occurred in maltose syrup at a relatively high concentration of ca. 20%. The process caused glucose concentration
to rise rapidly whereas accidentally more favourable weather conditions, earlier feeding of bees to
prepare them for wintering, exceptionally strong colonies and other factors coincided to accelerate
enzymatic hydrolysis of complex sugars thereby causing rapid crystallization of the stores. The bees
processed the fed syrup so thoroughly that it led to excessive glucose concentration and consequently, to glucose crystallization in comb cells.

Keywords: honey, starch syrup, wintering of bees, carbohydrates, crystallization, HPLC,

adulteration.

24

INTRODUCTION
Qualitative and quantitative assays of
saccharides in honey using HPLC or GC is
currently used on a wide scale to identify
honey surrogates and adulterations of
honey (Low and Sporns 1988, Swallow
and Low 1994, Low and South 1995,
Bogdanov 1999, Bogdanov and Martin 2002, Cotte et al. 2003) and to differentiate between some honey varieties
(Sabatini et al. 1989, 1990; Low et al.
1988; Ohe W. von der and Ohe K. von
d e r 1996; P e r s a n o O d d o and P i r o
2004; Cotte et al. 2004; Persano Oddo
and Bogdanov et al. 2004). Sugar analysis with the use of those methods is also becoming a routine test in this country
(Rybak-Chmielewska and Szczsna
2000; Ry b a k - C h m i e l e w s k a and
Szczsna 2003; Rybak-Chmielewska
and Konopacka 2005; Szczsna et al.
2003; Ry b a k - C h m i e l e w s k a et al.
2006a, 2006b). Sugar composition significantly different from that of honey (sugar
profile) allows the identification and detection of honey substitutes and surrogates
which arise without the involvement of
bees. On the other hand, a particularly difficult problem of honey being adulterated
with syrups following their processing by
bees has been solved only in part. White
and Doner (1978) while investigating natural carbon isotopes 13C and 12C in the
nectars of melliferous plants and in the sugars of maple syrup and of sugar cane demonstrated that the ratio of the two natural
isotopes was quite different in either case.
Consequently, it allows the detection of
honey adulteration with those syrups. It is
estimated that using that method, by assaying carbon isotopes, as small an addition of
syrups as 7% can be detected following the
processing the syrups by bees. Improved
(White and Winters 1989) and tested by
the team of the authors (White et al. 1998)
the method is however limited only to the

identification of sugar syrups derived from

sugar cane and from maize. It is also recommended by the Official Methods of
Analysis and by the Codex Alimentarius
Standard as one of the methods to detect
honey adulterations with high-fructose
sugar syrup (HFSS). Commonly used in
the laboratories throughout the world it
none-the-less fails to stand the test in the
case of honey adulteration with sugar
cane-derived sucrose (HFCS) done by
feeding bees with the solution of that sugar
and mixing it with honey. In the countries
in which commercial sugar (sucrose) processed from sugar beets is used the method
is unusable. Sucrose from that material has
a 13C to 12C ratio similar to that in sugars
of the nectars of melliferous plants. Because of that B o g d a n o v and M a r t i n
(2002) in their review of honey authenticity
studies recommend also other standard
methods such as specific electric conductivity tests to be used along with HPLC or
GC studies dealing with sugar profile
( Vo r w o h l 1964, P i a z z a et al. 1991,
Ry b a k and A c h r e m o w i c z 1986) and
proline content assays (D a v i e s 1975,
1978; White and Rudyj 1978; Rybak
and Achremowicz; Ohe von der et al.
1991) as well as the microscopic analysis
of honey sediment (Louveau et al. 1978,
Kerkvliet et al. 1995, Kerkvliet and
Meijer 2000, Ohe von der et al. 2004)
used most frequently as the so-called rough
screening methods which are aimed at the
detection of coarse adulterations. More
efficient methods are continued to be
searched for and possibilities provided by
new methods are tested e.g. quality parameters of honey are analyzed (including carbohydrates) using infrared spectroscopy:
near-infrared (NIR) spectroscopy (Garcia
Alvarez et al. 2002) and MIR spectroscopy to assay fructose, glucose, sucrose,
maltose and erlose and also assays of other
parameters (Lichtenberg-Kraag et al.
2002), phenolic acids and polyphenols in

Vol. 51 No. 1 2007

Journal of Apicultural Science

various honeys (A m i o t et al. 1989,

Tomas-Barberan et al. 2001) as well as
volatile compounds characteristic of different honey varieties (Ruoff 2003, Wolski
et al. 2006) mainly to develop new methods of classification and to complement the
description of unifloral honeys (Bogdanov
et al. 2004). Chosen and tested over
broader material, the methods to assay selected as the introduced and implemented
GC and HPLC techniques to examine sugars under the international honey standards
(Codex Alimentarius Commission Standard 2001, Council Directive 2001/110/EC
2002) will serve in the future the
beekeeping practice to confirm the unique
composition, high quality and purity of the
product. An analysis for the occurence of
starch dextrines, simple and yet effective in
restricting the presence of starch syrup, has
now been included in the Polish Standard
PN-88/A-77626 Mid pszczeli (Honeybee Honey). The dextrines usually arise in
small amounts during the manufacturing
process of those syrups (Horubaa 1975,
Tomasik 2000). The analysis included in
this countrys regulatory document in the
60s and used to detect adulteration of
honey with starch syrup, one of the traits
that disqualify honey, continues to be used
for that purpose. Along with chromatograph methods (F i o r i et al. 2000,
Rybak-Chmielew- ska et al. 2006b) it
can be successfully used to detect wheat
starch syrup in honey.
An extensive study of the sugars in syrups and honeys assayed with chromatograph techniques, GC and LC, was presented by the team of Cotte et al. (2003).
Based on the results obtained they determined the commercial syrups produced in
France to be detectable when added at a
rate of 5% to 10% to the investigated honeys of acacia, sweet chestnut and lavender. The authors determined such traits as
fructose and glucose contents, fructose to
glucose ratio as well as sucrose to maltose

25

ratio, maltotriose to total trisaccharides ratio, maltotriose to turanose ratio. While

analysing the syrups they observed that
generally their contents of fructose and glucose were lower than those in honeys. The
syrups had ca. 40 30% of sucrose or maltose. In a syrup with a high maltose content
the authors found a substantial maltotriose
content (6.5%). However, they did not investigate the sugar composition of the syrups once they were processed by bees.
The aim of this study was:
1. Assaying the following products for
carbohydrate contents:
a) syrup manufactured in Poland from
wheat starch which has been tried to
be used by beekeepers as bee food for
several years;
b) food processed from that syrup and
deposited by bees as winter stores;
c) samples of unifloral honeys of proven
quality and botanical origin;
2. Comparison of sugar compositions of
those products and an answer to the
questions:
a) To what extent does the HPLC
method to assay individual sugars aid
the identification of admixtures of
bee-inverted starch syrup in honey?
b) Why in the 2005/2006 season in some
of the apiaries did the starch syrup fed
to bees before winter undergo crystallization once it was deposited by bees
in the combs?

MATERIAL AND METHODS

Experiment material consisted of:
a) 6 samples of starch syrup produced
from wheat in Poland and composed
of: 3.3 4.1% fructose, ca. 22% glucose, 25.3 29.3% sucrose and
11.5% to 20.5% maltose. The syrup
with the contents of individual sugars
within the ranges as stated was fed to
8 bee colonies in one of the apiaries

26

Fig. 0. The percent participation of unifloral honey samples used for the study.
of the Bee Breeding Department,
Apiculture Division, Institute of
Pomology and Floriculture in Puawy
in the autumn of 2004.
b) 31 samples from the stores processed
by bees from the syrup deposited in
combs, 16 samples being collected
from the same apiary:
8 samples of winter store I - material collected from honeycombs of
bee colonies two months after
starch syrup was fed to bees in the
autumn;
8 samples of winter store II - material collected from the same bee
colonies following seven months of
feeding of bees (spring of 2005);
15 samples of winter store III samples of winter stores which
crystallized in honeycombs collected most of the time in early
spring of 2006 (sent in by
beekeepers from different regions
of Poland from the season of
2005/2006); samples collected towards the end of October of 2004
(store I).
c) 62 honey samples of confirmed botanical origin, and of verified
organoleptic and physico-chemical
features. The samples originated from

the apiaries of the Apiculture Division and from apiaries owned by

beekeepers who sent in honey samples for testing in 2005 (Fig. 0).
Research methods
Sugar content tests were performed using HPLC according to Bogdanov et al.
(1997) on a high-pressure SHIMADZU liquid chromatograph equipped with
LC-10ATVP liquid chromatograph pumps,
DGU-14A degasser, CTO-10AVP column
thermostat, RID-10A refractometric detector, POL-LAB CHROMA 2001 software
and SUPELCOSIL LC-NH2), 25 cm x
x 4.6 mm, 5 m chromatograph column.
The amount of the sample injected onto the
column was 20 l. The separation was conducted at a temperature of 30C with the
mobile phase acetonitrile : water (8:2) at a
flow rate of 1.3 ml/min. The identification
of sugars in honey was done by comparing
retention times of individual sugars in the
reference vs. tested solution (qualitative
analysis). The quantitative assays were
made of the following carbohydrates: fructose, glucose, sucrose, turanose, maltose,
isomaltose and trehalose. The contents of
those compounds were assayed based on
the comparing peak areas obtained in the
examined samples with those from the reference solution (quantitative analysis). To

Vol. 51 No. 1 2007

Journal of Apicultural Science

make the presentation of the obtained results more comprehensive the following
was calculated: total sugars, fructose to
glucose ratio and total monosaccharides.
The results concerning the contents of
sugars within the groups were processed
statistically by ANOVA. Duncans test at a
significance level of =0.05 was used to
examine the significance of differences between mean sugar contents in the treatments under comparison.

RESULTS AND DISCUSSION

In tables 1-10 and in Fig. 1-10 results
concerning the sugar contents of bee-processed starch syrup vs. those of honey were
compiled. Statistically significant differences in the sugar contents to make comparisons of winter store samples vs. honey
samples were obtained for isomaltose (Table 7) and for fructose to glucose ratio
(F/G) - Table 8, the latter being: 0.82 in
winter store I, 0.97 in winter store II and
0.50 in winter store III (nectar honey). The
lowest values of that trait within the honeys
were found for rape honey (0.98 Fig. 8).
The mean isomaltose contents for the respective store groups were: 0.48, 0.45 and
0.43%. The values were more than twice as
high as those for the nectar honey and
twice as low as those in honeydew honey
samples. However, because of generally
low isomaltose contents of the products under comparison and of the honeys it seems
that the content of that sugar cannot be a
good distinguisher for adulterated honey
made by bees from starch syrups (Table 7
and Fig. 7). More significant differences
were found for sugar contents of bee-processed starch syrups when they were compared with the contents of the same sugars
separately for nectar and honeydew honeys. Significant differences between the
compared treatments stores vs. nectar
honey were found for the following traits:
fructose content (Table 1), fructose to glu-

27

cose ratio (F/G Table 8), and for

disaccharides sucrose content (Table 3),
maltose content (Table 5) and isomaltose
content (Table 7) and also for total
monosaccharides (Table 9). Earlier on,
Ohe von der and Schnberger (2002),
and Liebig (2005) while discussing the
composition of starch syrups and their suitability in beekeeping paid attention to substantial contents of maltose which were retained also in bee-processed winter stores
deposited in the combs. The investigators
also made note of the fact that the maltose
content of the stores gradually decreased
with the concomitant increase in glucose
content. It was also confirmed by the results of this study. Comparison of the sugar
content of the stores collected and tested
following 2 months (store I) and 7 months
(store II) after feeding sugar syrup to bees
gave evidence that average maltose content
decreased from 6.64% to 5.48% (Table 5)
and glucose content showed a slight increase from 31.05% to 31.59% (Table 2).
When samples of starch syrup samples
were compared with honeydew honey samples the two products differed only for two
traits: greater amounts of the disaccharides
turanose and trehalose were found in honeydew honey. The difference was true not
only of the comparison of honeydew honey
vs. stores but also of stores vs. nectar honey
(Tables 4 and 6 and Fig. 4 and 6). It is a
characteristic feature of honeydew honeys.
The sugar content values for fructose
varied significantly among the store groups
tested. In the samples collected for testing
two months after feeding (store I) the fructose content averaged 24.89% whereas in
the samples collected 7 months after the
syrup was fed to bees (store II) the average
content of that simple sugar was 30.71%.
Likewise, the fructose to glucose ratio was
lower in stores samples collected 2 months
after the feeding (store I) date averaging
0.82 (Table 8). However, after 7 months it
reached a value characteristic of rape

28
honey (Table 8 and Fig. 8). With the passage of time, from the date on which the
syrup was fed to bees onwards, the content
of the disaccharide sucrose significantly
declined from an average of 5.66% to
2.30% (Table 3). It is readily evident even
now that store II (centrifuged in the spring,
seven months after the bees were fed)
meets the requirements as to carbohydrate
contents set down by the regulatory documents concerning honey quality currently
in force. According to those requirements
sucrose content must not be higher than 5%

and total saccharides content must not be

lower than 60% for nectar honeys and 40%
for honeydew honeys. The average content
of fructose (30.71% - Table 1) and of glucose (31.59% - Table 2) gives a product
that contains 62.3% monosaccharides so
that it also conforms with the standing requirement for honey. In Table 9 and in
Fig. 9 the compiled results for that parameter average 71.74% for nectar honeys and
65.39% for honeydew honeys. Thus it
seems justified to verify those requirements

Table 1
Fructose content of winter stores made from starch syrup by the bees vs. that of nectar and
honeydew honeys.
Studied material

Fructose content %
Mean

From

To

3.62

3.30

4.10

Winter store I1)

24.89 b4)

21.35

30.85

Winter store II2)

30.71 c

28.67

33.09

Winter store III3)

19.73 a

12.50

26.40

Nectar honey

38.81 d

38.47

39.63

Honeydew honey

35.31 cd

34.90

35.95

Starch syrup

1)

Winter store I - material collected from honeycombs of bee colonies two months
after starch syrup was fed to bees in the autumn;
2) Winter store II - material collected from the same bee colonies following seven months
of feeding of bees (spring of 2005);
3) Winter store III - samples of winter stores which crystallized in honeycombs collected most
of the time in early spring of 2006 (sent in by beekeepers from different regions of Poland
from the season of 2005/2006);
4) abcd - differences statistically significant between means in rows at =0.05.

Fig. 1. Fructose content of unifloral honeys vs. that of starch syrup and winter stores made
from that syrup by the bees (mean values).

Vol. 51 No. 1 2007

Journal of Apicultural Science

29

Table 2
Glucose content of winter stores made from starch syrup by the bees vs. that of nectar and
honeydew honeys.
Studied material

Glucose content %
Mean

From

To

21.95

21.10

22.40

Winter store I1)*

31.05 a4)

29.18

33.07

Winter store II2)

31.59 a

29.83

32.88

Winter store III3)

38.98 b

33.50

41.60

Nectar honey

32.90 a

32.42

33.17

Honeydew honey

30.08 a

29.40

31.00

Starch syrup

For explanation: see Table 1.

Fig. 2. Glucose content of unifloral honeys vs. that of starch syrup and winter stores made
from that syrup by the bees (mean values).

Table 3
Sucrose content of winter stores made from starch syrup by the bees vs. that of nectar and
honeydew honeys.
Studied material
Starch syrup

Sucrose content %
Mean

From

To

27.32

25.30

29.30

4.31

7.17

Winter store

I1)*

Winter store

II2)

2.30 b

0.92

3.88

Winter store

III3)

2.49 b

0.85

3.50

Nectar honey

0.49 a

0.17

0.63

Honeydew honey

1.26 ab

0.85

1.65

For explanation: see Table 1.

5.66

c4)

30

Fig. 3. Sucrose content of unifloral honeys vs. that of starch syrup and winter stores made
from that syrup by the bees (mean values).

Table 4
Turanose content of winter stores made from starch syrup by the bees vs. that of nectar and
honeydew honeys.
Studied material

Turanose content %
Mean

From

To

0.00

0.00

0.00

Winter store I1)*

0.98 a4)

0.63

1.44

Winter store II2)

1.44 b

1.31

1.63

Winter store III3)

0.81 a

0.50

1.30

Nectar honey

1.15 ab

1.02

1.31

1.83 c

1.70

2.05

Starch syrup

Honeydew honey
*

For explanation: see Table 1.

Fig. 4. Turanose content of unifloral honeys vs. that of starch syrup and winter stores made
from that syrup by the bees (mean values).

Vol. 51 No. 1 2007

Journal of Apicultural Science

31

Table 5
Maltose content of winter stores made from starch syrup by the bees vs. that of nectar and
honeydew honeys.
Studied material

Maltose content %
Mean

From

To

17.42

11.50

20.50

Winter store I1)*

6.64 cd4)

5.39

7.22

Winter store II2)

5.48 bc

4.55

6.20

Winter store III3)

8.50 d

5.21

11.80

Nectar honey

2.33 a

2.03

2.68

Honeydew honey

3.61 ab

3.35

3.90

Starch syrup

For explanation: see Table 1.

Fig. 5. Maltose content of unifloral honeys vs. that of starch syrup and winter stores made
from that syrup by the bees (mean values).

Table 6
Trehalose content of winter stores made from starch syrup by the bees vs. that of nectar and
honeydew honeys.
Studied material
Starch syrup
Winter store

I1)*

Trehalose content %
Mean

From

To

0.00

0.00

0.00

0.30

0.51

0.41

a4)

Winter store II2)

0.66 a

0.62

0.72

Winter store III3)

0.43 a

0.30

0.70

Nectar honey

0.70 a

0.52

0.87

Honeydew honey

1.71 b

1.25

2.00

For explanation: see Table 1.

32

Fig. 6. Trehalose content of unifloral honeys vs. that of starch syrup and winter stores made
from that syrup by the bees (mean values).

Table 7
Isomaltose content of winter stores made from starch syrup by the bees vs. that of nectar
and honeydew honeys.
Studied material
Starch syrup

Isomaltose content %
Mean

From

To

0.00

0.00

0.00

0.44

0.51

Winter store

I1)*

Winter store

II2)

0.45 b

0.42

0.50

Winter store III3)

0.43 b

0.30

0.70

Nectar honey

0.20 a

0.13

0.25

Honeydew honey

0.88 c

0.83

0.95

0.48

b4)

For explanation: see Table 1.

Fig. 7. Isomaltose content of unifloral honeys vs. that of starch syrup and winter stores made
from that syrup by the bees (mean values).

Vol. 51 No. 1 2007

Journal of Apicultural Science

33

Table 8
Fructose/Glucose (F/G) ratio of winter stores made from starch syrup by the bees
vs. that of nectar and honeydew honeys.
Studied material

F/G
Mean

From

To

0.16

0.15

0.17

Winter store I1)*

0.82 b4)

0.68

1.05

Winter store II2)

0.97 c

0.89

1.05

Winter store III3)

0.50 a

0.37

0.65

Nectar honey

1.19 d

1.17

1.21

Honeydew honey

1.18 d

1.16

1.20

Starch syrup

For explanation: see Table 1.

Fig. 8. Fructose/Glucose (F/G) ratio of unifloral honeys vs. that of starch syrup and winter
stores made from that syrup by the bees (mean values).

Table 9
Monosaccharides content of winter stores made from starch syrup by the bees
vs. that of nectar and honeydew honeys.

Studied material
Starch syrup

Monosaccharides content %
Mean

From

To

25.60

24.70

26.30

I1)*

55.93a4)

52.83

60.03

Winter store II2)

62.04ab

59.71

64.63

Winter store III3)

58.70ab

46.00

67.10

Nectar honey

71.74 c

70.88

72.8

Honeydew honey

65.39 bc

64.60

66.95

Winter store

For explanation: see Table 1.

34

Fig. 9. Monosaccharides content of unifloral honeys vs. that of starch syrup and winter stores
made from that syrup by the bees (mean values).

Table 10
Total sugar content of winter stores made from starch syrup by the bees
vs. that of nectar and honeydew honeys.
Studied material

Saccharides content %
Mean

From

To

70.94

64.60

75.20

Winter store I1)*

72.24 a4)

70.70

74.78

Winter store II2)

75.24 b

74.52

76.60

III3)

74.80 b

71.50

76.70

Nectar honey

76.92 b

75.96

77.49

Honeydew honey

76.03 b

74.05

77.55

Starch syrup

Winter store

For explanation: see Table 1.

Fig. 10. Total sugar content of unifloral honeys vs. that of starch syrup and winter stores made
from that syrup by the bees (mean values).

Vol. 51 No. 1 2007

Journal of Apicultural Science

when the domestic regulatory documents

are revised.
The investigations of the spectra of sugars in syrups and honeys by means of assays using two chromatograph techniques GC and LC were also presented by the
team of C o t t e et al. (2003) of France.
Based on the results obtained they stated
that as small an addition of commercial
syrups to honey as 5 10% is already detectable. The investigators singled out the
following most suitable traits that distinguish syrups from honey: fructose content,
glucose content, fructose to glucose ratio,
and also the ratios of sucrose to malotose,
maltotoriose to total trisaccharides and
maltotriose to turanose. Based on the results of this study the same traits would be
chosen, although without such trisaccharides as maltotriose and others because the
HPLC method with refractometer detector
and the column selected for this study did
not allow good separation of sugars within
that range nor did they permit their accurate quantitative determination. However,
the team of Cotte did not investigate the
sugar composition of syrups once they
were processed by bees.
The problem of the crystallization of
stores following their deposition in the
combs by bees as winter food requires separate treatment. The crystallization occurred
only in a part of apiaries that were fed starch
syrup as winter store in the season of
2005/2006. The analysis of the content of
individual sugars in the samples of those
stores (store III) aided the understanding and
the explanation of the problem. It turned out
that in that group of samples the content of
fructose was much lower and the content of
glucose was higher. It caused that sugar to
crystallize in the comb cells. According to
Ohe von der and Schnberger (2000)
the critical solution saturation point with
glucose is 32 g/100 g and above that concentration glucose crystallization occurs. In
the tested samples (store III) the glucose
concentration was on average as high as
38.98% (Table 2).

35

CONCLUSIONS
1. Lower than in honey fructose content
(below 32%), high (over 5%) maltose
content and a low F/G ratio averaging
0.76 vs. 1.18 in honey samples may be
used as a distinguisher to identify
products processed by bees from
starch syrups.
2. The store centrifuged in the spring
(seven months after the syrup was fed
to bees) reached a carbohydrate content in conformance with the requirements in the valid regulatory documents concerning honey quality. A more
in depth analysis of those requirements
seems to be required while revising the
current standard PN-88/A-77626 Mid
pszczeli (Honeybee Honey).
3. In the apiaries in which the crystallization of the starch syrup-derived store
occurred the conditions had arisen
which were more propitious for the
hydrolysis of complex sugars mainly
maltose to glucose such as favourable
weather conditions, earlier feeding of
bees exceptionally strong bee colonies.
Previously, such conditions favoured
good wintering of bee colonies which
were fed sugar syrup (sucrose solution) but it was not true of this case. It
was because the bees broke down the
maltose-enriched syrup so thouroughly
that it led to a considerable buildup of
glucose and its consequent crystallization. In those samples glucose concentration was found to average 38.98%.

REFERENCES
Amiot M.J. Aubert S. Gonnet M.
Tacchini M. (1989) Phenolic composition of honeys: preliminary study on identyfication and group quantification. Apidologie
20: 115-125.
Bogdanov S. (1999)
Honey
quality,
methods of analysis and international
regulatory standards: review of the work of
the International Honey Commission. Mitt.
Gebiete Lebensm. Hyg., 90:108-125.

36
Bogdanov S., Martin, P. (2002) Honey
authenticity: a review. Mitt. Gebiete
Lebensm. Hyg., 93: 232-254.

Kerkvliet J.D., Meijer H.A.J. (2000)

Adulteration of honey: relation between
microscopic analysis and 13C measurements,
Apidologie 31, 717-726.

Bogdanov S., Martin P., Lllmann C.

(1997) Harmonised methods of the
European Honey Commission. Apidologie
(extra issue): 1-59.

Liebig G. (2005)
Getreidestrkesirup:
besser als sein Ruf. Deutsches Binen Journal
13(8).

Bogdanov S., Ruoff K., Persano-Oddo

L. (2004) Physico-chemical methods for
the characterization of unifloral honeys: a
review. Apidologie 35: S4-S17.

Lichtenberg-Kraag B., Hedtke C.,

Bienefeld K. (2002) Infrared spectroscopy in routine quality analysis of honey.
Apidologie, 33: 327-337.

Codex
Alimentarius
Commission
Standard (2001) 24th Session, July
2001, adopting the draft revised standard for
honey. Alinorm 01/25, Appendix II: 22-24.

Louveaux J., Maurizio A. Vorwohl G.

(1978) Methods of melissopalynology.
Bee World, 59: 139-162.

Cotte J.F., Casabianca H., Chardon S.,

Lheritier J., Grenier-Loustalot M.F.
(2003) Application of carbohydrate
analysis to verify honey authenticity. J.
Chromatogr. A. 1021: 145-155.
Cotte J.F., Casabianca H., Chardon S.,
Lheritier J., Grenier-Loustalot M. F.
(2004) Chromatographic analysis of
sugars applied to the characterization of
monofloral honey. Anal Bioanal Chem 308:
698-705.
Council Directive 2001/110/EC of 20
December 2001 relating to honey (2002)
Official Journal of the European
Communities, L 10: 47 - 52.
Davies A.M. (1975) Amino acid analysis
of honeys from eleven countries. J. apic.
Res., 14(1): 29 - 39.
Fiori J., Serra G., Sabatini A.G., Zucchi
P., Barbattini R., Gazziola F. (2000)
Dextrins HPLC analysis in Matcalfa
pruinosa (Say) honeydew, Ind. Aliment. 39:
463-466.
Garcia Alvarez M., Ceresuela S.,
Huidobro j.f. Hermida M., Rodriguez
Otero J.L. (2002) Determination of
polarimetric parameters of honey by near-infrared transflectance spectroscopy, J.
Agric. Food Chem., 50. 419-425.
Horubaa A. (1975) Podstawy Przechowalnictwa ywnoci. PWN Warszawa 417.
Kerkvliet J.D., Shrestha M., Tuladhar
K., Manandhar H. (1995) Microscopic
detection of adulteration of honey with cane
sugar and cane sugar products. Apidologie
26: 131-139.

Low N.H., Sporns P. (1988)

Analysis
and quantification of minor, di- and
trisaccharides in honey, using capillary gas
chromatography. J. Food Sci., 53: 558-561.
Low N.H., Nelson D.L., Sporns P.
(1988) Carbohydrate analysis of western
Canadian honeys and their nectar sources to
determinethe
origin
of
honey
oligosaccharides, J. apic. Res., 27, 245-251.
Low N.H., South W. (1995)
Determination of honey authenticity be
capillary gas chromatography. J. AOAC Int.,
78(5): 1210-1218.
Ohe W.von der, Ohe K.von der (1996)
Characterisation of honeydew honey with
specific saccharides. Apidologie, 27:
270-272.
Ohe W. von der, Schnberger H. (2000)
Fr die Ernhrung der Bienen: Futtersirup
im Vergleich. Deutsches Bienen-Journal
8(8): 312-314.
Ohe W. von der, Schnberger H. (2002)
Bienenernahrung: Futtersirup im Vergleich.
Binenvater, 123(9): 11-15.
Ohe W. von der, Dustmann J.H., Von der
Ohe K. (1991) Prolin als Kriterium der
Reife des Honigs. Dtsch. Lebensm.
Rundsch., 87: 383-386.
Ohe W. von der, Persano Oddo L., Piana
M.L., Marlot M., Martin P. (2004)
Harmonized methods of melissopalynology,
Apidologie (Suppl. 1), 18-25.
Persano Oddo L., Piro R. (2004) Main
European unifloral honeys: descriptive
sheets. Apidologie 35: 38-81.

Vol. 51 No. 1 2007

Journal of Apicultural Science

Persano Oddo L., Bogdanov S. (2004)

Determination of honey botanical origin:
problems and issues Apidologie 35: S2-S3.
Piazza M. G.,Accorti M.,Persano Oddo
L. (1991) Electrical conductivity, ash,
colour and specific rotatory power in Italian
unifloral honeys. Apicoltura, 7: 63.
Polska Norma PN-88/A-77626 Mid
pszczeli, 1998. Dziennik Norm i Miar nr 8,
1998. Wydawnictwa Normalizacyjne Alfa.
Ruoff, K. (2003) Solid-phase microextraction of honey volatiles: a method for
the determination of the botanical origin of
honey, Master Thesis, University of
Helsinki.
Rybak H., Achremowicz B. (1986)
Zmiany w skadzie chemicznym miodw
naturalnych i zafaszowanych zinwertowan
przez pszczoy sacharoz zachodzce
podczas przechowywania. Pszczeln. Zesz.
Nauk., 30: 19-35.
Rybak-Chmielewska H., Szczsna T.
(2000) Mono- and oligosaccharides
composition of some Polish unifloral honeys
by means of gas chromatography. Pszczeln.
Zesz. Nauk., 44(2): 325-334.

37

Sabatini A.G., Persano Oddo L., Piazza

M.G., Accorti M., Marcazzan G. (1990)
Glucide spectrum in the Italian unifloral
honeys. II. Di-and trisaccharides. Apicoltura,
6: 63-67.
Szczsna T., Rybak-Chmielewska H.,
Skubida P. (2003) Contribution to the
understanding of the phenomenon of cement
honey. J. apic. Sci., 47(2): 103-108.
Swallow K. W., Low N.H. (1994)
Determination of honey authenticity by
anion-exchange liquid chromatography. J.
AOAC Int., 77(3): 695-702.
Tomas-Barberan F.A., Martos I.,
Ferreres F., Radovic B.S., Anklam E.
(2001) HPLC flavonoid profiles as
markers fort he botanical origin of European
unifloral honeys, J. Sci. Food Agric., 81:
485-496
Tomasik P. (2000)
Waciwoci
funkcjonalne sacharydw. Rozdzia 6 W:
Chemia ywnoci pod redakcj Sikorskiego
Z.E. wydanie III, Wydawnictwo Naukowo-Techniczne, Warszawa: 164-184.

Rybak-Chmielewska H., Szczsna T.

(2003) Determination of saccharides in
multifloral honey by means of HPLC. J.
apic. Sci., 47(2): 93-101.

Vorwohl G. (1964) Die Messung der

elektrischen Leitfahigkeit des Honigs und
die Verwendung der Messwerte zur
Sortendiagnose und zum Nachweis von
Verfalschungen mit Zuckerfutterungshonig.
Z. f. Bienenforsch., 7: 37-47.

Rybak-Chmielewska H., Konopacka Z.

(2005)
Co
to
jest
izoglukoza.
Pszczelarstwo 6: 6-7.

White J.W. Jr., Doner L.W. (1978) The

13C/12C ratio in honey. J. apic. Res., 17(3):
94-99.

Rybak-Chmielewska H., Szczsna T.,

Biekowska M. (2006a)
Gas
chromatography (GC) study of sugar
composition in honeys and winter stores
processed by bees from sucrose syrups. J.
apic. Sci., 50(2): 147-155.

White J.W. Jr., Rudyj O.N. (1978)

Proline content of United States honeys. J.
apic. Res., 17(2): 89-93.

Rybak-Chmielewska H., Szczsna T.,

Wa E. (2006b) Attempt to assay
maltodextrins occurring in starch syrup and
in winter stores made by bees from that
syrups. J. apic. Sci., 50(2): 127-135.
Sabatini A.G., Persano Oddo L., Piazza
M.G., Accorti M., Marcazzan G. (1989)
Glucide spectrum in the Italian unifloral
honeys. I. Fructose and glucose. Apicoltura,
5: 35-46.

White J.W., Winters K. (1989) Honey

protein as internal standard for stable isotope
ratio detection of adulteration of honey. J.
Ass. Off. Anal. Chem., 72: 907-911.
White J.W., Winters K., Martin P.,
Rossmann A. (1998) Stable carbon
isotope ratio analysis of honey: Validation of
internal standard procedure for worldwide
application. J. Ass. Off. Anal. Chem. 81:
610-619.
Wolski T., Tambor K., Rybak-Chmielewska H., Kdzia B. (2006)
Identification of honey volatile components
by solid phase microextraction (SPME) and
gas chromatography/mass spectrometry
(GC/MS). J. apic. Sci., 50(2): 115-126.

38

BADANIA CHROMATOGRAFICZNE (HPLC) SKADU

CUKRW W MIODACH I ZAPASACH NA ZIM
WYTWORZONYCH PRZEZ PSZCZOY Z SYROPW
SKROBIOWYCH
Rybak-Chmielewska

H.

S t r e s z c z e n i e
Okrelono skad cukrw w trzech grupach produktw: w syropie skrobiowym (maltozowym)
produkowanym w Polsce ze skrobi pszenicy (ktry od kilku lat pszczelarze prbuj
wykorzystywa jako pokarm dla pszcz); w wytworzonym z tego syropu przez pszczoy
pokarmie stanowicym zapas na zim oraz w miodzie. Zapasy zimowe analizowano w trzech
podgrupach: podgrupa I - materia pobrany z plastrw rodzin pszczelich po dwch miesicach
od jesiennego podkarmiania pszcz syropem (koniec padziernika 2004); podgrupa II
materia pobrany z tych samych rodzin pszczelich po siedmiu miesicach od podkarmiania
(wiosn 2005 roku) i podgrupa III - prbki zapasu zimowego, ktry skrystalizowa w plastrach,
pobierany najczciej wczesn wiosn 2006 (przysyany przez pszczelarzy z rnych rejonw
Polski z sezonu 2005/2006).
Badania zawartoci cukrw wykonano metod HPLC z detektorem refraktometrycznym wg
B o g d a n o v a i in. (1997). Oznaczono i porwnano zawartoci nastpujcych cukrw: glukozy,
fruktozy, sacharozy, maltozy, izomaltozy, turanozy i trehalozy.
W porwnywanych wynikach zawartoci poszczeglnych cukrw w zapasach utworzonych
z syropu i w miodzie odnaleziono istotne rnice. Rutynowe badania skadu cukrw metod
HPLC mog by pomocne przy identyfikacji produktw wytworzonych przez pszczoy
z syropw maltozowych. Wyrnikiem dla tych produktw bya nisza w stosunku do miodu
zawarto fruktozy (poniej 32%); wysoka (ponad 5%) zawarto maltozy oraz niski stosunek
zawartoci fruktozy do glukozy (F/G), rednia z badanych podgrup 0,76, podczas gdy
w prbkach miodw warto ta wynosia rednio 1,18. Najnisze wartoci tego parametru dla
miodw charakteryzoway mid rzepakowy i wynosiy rednio 0,98.
Zosta te wyjaniony problem krystalizacji zapasw z syropu maltozowego po zoeniu ich
przez pszczoy w plastrach jako pokarmu na zim. Krystalizacja wystpia tylko w czci pasiek
karmionych na zim 2005/2006 syropem maltozowym. Okazao si, e w tej grupie prbek w
stosunku do pozostaych (nieskrystalizowanych lub tylko czciowo skrystalizowanych)
zawarto fruktozy bya istotnie nisza, natomiast wysza zawarto glukozy. Wg O h e v o n
d e r i S c h n b e r g e r a (2000) punkt krytyczny nasycenia roztworu glukoz wynosi 32 g/100 g,
powyej tego stenia nastpuje jej krystalizacja. W omawianych prbkach zapasw stenie
glukozy wynosio rednio a 38,98%. Spowodowao to krystalizacj tego cukru ju
w komrkach plastrw. W pasiekach, w ktrych nastpia krystalizacja zapasu z syropu
skrobiowego (maltozowego) zaistniay bardziej sprzyjajce warunki enzymatycznej hydrolizy
cukrw zoonych do cukrw prostych. Szczeglne znaczenie ma tu rozkad maltozy, ktrej
w tym syropie byo stosunkowo duo okoo 20%. Proces ten powodowa szybki wzrost
stenia glukozy, a losowo bardziej korzystne warunki pogodowe, wczeniejsze karmienie
pszcz na zim, wyjtkowo silne rodziny i inne czynniki, przypieszajc proces enzymatycznej
hydrolizy cukrw zoonych, spowodoway szybk krystalizacj wytworzonych zapasw.
Pszczoy tak dokadnie przetworzyy podany im syrop, e doprowadzio to do zbyt duego
stenia glukozy i w konsekwencji do jej krystalizacji w komrkach plastrw.

Sowa kluczowe: mid, syrop skrobiowy, zimowanie pszcz, wglowodany, krystalizacja,

HPLC, faszowanie.