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HIGH PERFORMANCE LIQUID CHROMATOGRAPHY(HPLC) STUDY OF SUGAR COMPOSITION IN SOMEKINDS OF NATURAL HONEY AND WINTER STORES

PROCESSED BY BEES FROM STARCH SYRUP

H e l e n a R y b a k - C h m i e l e w s k a

Research Institute of Pomology and Floriculture, Apiculture Division, Department of Bee Products,24-100 Pu³awy, Kazimierska 2, Poland. E-mail: [email protected]

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 producedin 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 sub-groups: subgroup 1 – material collected from honeycombs of bee colonies two months after thesyrup was fed to the bees in autumn (end of October of 2004); subgroup 2 – material collected fromthe same bee colonies following seven months of feeding (spring of 2005); subgroup 3 – samples ofwinter 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 toBogdanov et al. (1997).

The following sugars were assayed and compared: glucose, fructose, sucrose, maltose, isomal-tose, turanose and trehalose.

Significant differences were found for the contents of individual sugars in the stores made fromsyrup vs. those in the honey. Routine HPLC assays of sugars can be helpful in the identification ofproducts made by bees from maltose syrups. The main distinguisher for those products was fructosecontent (lower than that in honey by 32%), high maltose content (over 5%) and low fructose to glu-cose (F/G) ratio (0.76 when averaged across subgroups) whereas in honey samples it was 1.18 onaverage. 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 frommaltose syrup following their depositing in the combs as winter storage. The crystallization oc-curred only in part of the apiaries that were fed maltose syrup in the season of 2005/2006. It wasfound in that group of samples that fructose content was significantly lower and that of glucose sig-nificantly higher than in the remaining ones (non-crystallized or partly crystallized). According toOhe von der and Schönberger (2000) the critical point for at which a solution becomes satu-rated with glucose is 32 g/100 g and once that concentration is exceeded crystallization occurs. Inthe examined samples the concentration of glucose averaged as much as 38.98%. It caused the sugarto crystallize already in the comb cells. In the apiaries in which crystallization of maltose syrup de-rived stores occurred there were conditions which favoured enzymatic hydrolysis of complex sugarsto simple ones. Of particular importance here is the breakdown of maltose which occurred in mal-tose syrup at a relatively high concentration of ca. 20%. The process caused glucose concentrationto rise rapidly whereas accidentally more favourable weather conditions, earlier feeding of bees toprepare them for wintering, exceptionally strong colonies and other factors coincided to accelerateenzymatic hydrolysis of complex sugars thereby causing rapid crystallization of the stores. The beesprocessed the fed syrup so thoroughly that it led to excessive glucose concentration and conse-quently, to glucose crystallization in comb cells.

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

Vol. 51 No. 1 2007 Journal of Apicultural Science 23

INTRODUCTIONQualitative and quantitative assays of

saccharides in honey using HPLC or GC iscurrently used on a wide scale to identifyhoney surrogates and adulterations ofhoney (Low and Sporns 1988, Swallowand Low 1994, Low and South 1995,Bogdanov 1999, Bogdanov and Mar-tin 2002, Cotte et al. 2003) and to differ-entiate between some honey varieties(Sabatini et al. 1989, 1990; Low et al.1988; Ohe W. von der and Ohe K. vond e r 1996; P e r s a n o O d d o and P i r o2004; Cotte et al. 2004; Persano Oddoand Bogdanov et al. 2004). Sugar analy-sis with the use of those methods is also be-coming a routine test in this country(Rybak-Chmielewska and Szczêsna2000; Ry b a k - C h m i e l e w s k a andSzczêsna 2003; Rybak-Chmielewskaand Konopacka 2005; Szczêsna et al.2003; Ry b a k - C h m i e l e w s k a et al.2006a, 2006b). Sugar composition signifi-cantly different from that of honey (sugarprofile) allows the identification and detec-tion of honey substitutes and surrogateswhich arise without the involvement ofbees. On the other hand, a particularly diffi-cult problem of honey being adulteratedwith syrups following their processing bybees has been solved only in part. Whiteand Doner (1978) while investigating nat-ural carbon isotopes 13C and 12C in thenectars of melliferous plants and in the sug-ars of maple syrup and of sugar cane dem-onstrated that the ratio of the two naturalisotopes was quite different in either case.Consequently, it allows the detection ofhoney adulteration with those syrups. It isestimated that using that method, by assay-ing carbon isotopes, as small an addition ofsyrups as 7% can be detected following theprocessing the syrups by bees. Improved(White and Winters 1989) and tested bythe team of the authors (White et al. 1998)the method is however limited only to the

identification of sugar syrups derived fromsugar cane and from maize. It is also rec-ommended by the Official Methods ofAnalysis and by the Codex AlimentariusStandard as one of the methods to detecthoney adulterations with high-fructosesugar syrup (HFSS). Commonly used inthe laboratories throughout the world itnone-the-less fails to stand the test in thecase of honey adulteration with sugarcane-derived sucrose (HFCS) done byfeeding bees with the solution of that sugarand mixing it with honey. In the countriesin which commercial sugar (sucrose) pro-cessed from sugar beets is used the methodis unusable. Sucrose from that material hasa 13C to 12C ratio similar to that in sugarsof the nectars of melliferous plants. Be-cause of that Bogdanov and Mar t in(2002) in their review of honey authenticitystudies recommend also other standardmethods such as specific electric conduc-tivity tests to be used along with HPLC orGC studies dealing with sugar profile(Vorwoh l 1964, P i azza et al. 1991,Rybak and Achremowicz 1986) andproline content assays (Dav ie s 1975,1978; White and Rudyj 1978; Rybakand Achremowicz; Ohe von der et al.1991) as well as the microscopic analysisof honey sediment (Louveau et al. 1978,Kerkvliet et al. 1995, Kerkvliet andMeijer 2000, Ohe von der et al. 2004)used most frequently as the so-called roughscreening methods which are aimed at thedetection of “coarse” adulterations. Moreefficient methods are continued to besearched for and possibilities provided bynew methods are tested e.g. quality param-eters of honey are analyzed (including car-bohydrates) using infrared spectroscopy:near-infrared (NIR) spectroscopy (GarciaAlvarez et al. 2002) and MIR spectros-copy – to assay fructose, glucose, sucrose,maltose and erlose and also assays of otherparameters (Lichtenberg-Kraag et al.2002), phenolic acids and polyphenols in

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various honeys (A m i o t et al. 1989,Tomas-Barberan et al. 2001) as well asvolatile compounds characteristic of differ-ent honey varieties (Ruoff 2003, Wolskiet al. 2006) mainly to develop new meth-ods of classification and to complement thedescription of unifloral honeys (Bogdanovet al. 2004). Chosen and tested overbroader material, the methods to assay se-lected as the introduced and implementedGC and HPLC techniques to examine sug-ars under the international honey standards(Codex Alimentarius Commission Stan-dard 2001, Council Directive 2001/110/EC2002) will serve in the future thebeekeeping practice to confirm the uniquecomposition, high quality and purity of theproduct. An analysis for the occurence ofstarch dextrines, simple and yet effective inrestricting the presence of starch syrup, hasnow been included in the Polish StandardPN-88/A-77626 „Miód pszczeli” (Honey-bee Honey). The dextrines usually arise insmall amounts during the manufacturingprocess of those syrups (Horuba³a 1975,Tomasik 2000). The analysis included inthis country’s regulatory document in the60’s and used to detect adulteration ofhoney with starch syrup, one of the traitsthat disqualify honey, continues to be usedfor that purpose. Along with chroma-tograph methods (F i o r i et al. 2000,Rybak-Chmielew- ska et al. 2006b) itcan be successfully used to detect wheatstarch syrup in honey.

An extensive study of the sugars in syr-ups and honeys assayed with chromato-graph techniques, GC and LC, was pre-sented by the team of Cotte et al. (2003).Based on the results obtained they deter-mined the commercial syrups produced inFrance to be detectable when added at arate of 5% to 10% to the investigated ho-neys of acacia, sweet chestnut and laven-der. The authors determined such traits asfructose and glucose contents, fructose toglucose ratio as well as sucrose to maltose

ratio, maltotriose to total trisaccharides ra-tio, maltotriose to turanose ratio. Whileanalysing the syrups they observed thatgenerally their contents of fructose and glu-cose were lower than those in honeys. Thesyrups had ca. 40 – 30% of sucrose or malt-ose. In a syrup with a high maltose contentthe authors found a substantial maltotriosecontent (6.5%). However, they did not in-vestigate the sugar composition of the syr-ups once they were processed by bees.The aim of this study was:

1. Assaying the following products forcarbohydrate contents:

a) syrup manufactured in Poland fromwheat starch which has been tried tobe used by beekeepers as bee food forseveral years;

b) food processed from that syrup anddeposited by bees as winter stores;

c) samples of unifloral honeys of provenquality and botanical origin;

2. Comparison of sugar compositions ofthose products and an answer to thequestions:

a) To what extent does the HPLCmethod to assay individual sugars aidthe identification of admixtures ofbee-inverted starch syrup in honey?

b) Why in the 2005/2006 season in someof the apiaries did the starch syrup fedto bees before winter undergo crystal-lization once it was deposited by beesin the combs?

MATERIAL AND METHODSExperiment material consisted of:a) 6 samples of starch syrup produced

from wheat in Poland and composedof: 3.3 – 4.1% fructose, ca. 22% glu-cose, 25.3 – 29.3% sucrose and11.5% to 20.5% maltose. The syrupwith the contents of individual sugarswithin the ranges as stated was fed to8 bee colonies in one of the apiaries


of the Bee Breeding Department,Apiculture Division, Institute ofPomology and Floriculture in Pu³awyin the autumn of 2004.

b) 31 samples from the stores processedby bees from the syrup deposited incombs, 16 samples being collectedfrom the same apiary:

— 8 samples of winter store I - mate-rial collected from honeycombs ofbee colonies two months afterstarch syrup was fed to bees in theautumn;

— 8 samples of winter store II - mate-rial collected from the same beecolonies following seven months offeeding of bees (spring of 2005);

— 15 samples of winter store III -samples of winter stores whichcrystallized in honeycombs col-lected most of the time in earlyspring of 2006 (sent in bybeekeepers from different regionsof Poland from the season of2005/2006); samples collected to-wards the end of October of 2004(store I).

c) 62 honey samples of confirmed bo-tanical origin, and of verifiedorganoleptic and physico-chemicalfeatures. The samples originated from

the apiaries of the Apiculture Divi-sion and from apiaries owned bybeekeepers who sent in honey sam-ples for testing in 2005 (Fig. 0).

Research methodsSugar content tests were performed us-

ing HPLC according to Bogdanov et al.(1997) on a high-pressure SHIMADZU liq-uid chromatograph equipped withLC-10ATVP liquid chromatograph pumps,DGU-14A degasser, CTO-10AVP columnthermostat, RID-10A refractometric detec-tor, POL-LAB CHROMA 2001 softwareand SUPELCOSIL LC-NH2), 25 cm xx 4.6 mm, 5 µm chromatograph column.The amount of the sample injected onto thecolumn was 20 µl. The separation was con-ducted at a temperature of 30°C with themobile phase acetonitrile : water (8:2) at aflow rate of 1.3 ml/min. The identificationof sugars in honey was done by comparingretention times of individual sugars in thereference vs. tested solution (qualitativeanalysis). The quantitative assays weremade of the following carbohydrates: fruc-tose, glucose, sucrose, turanose, maltose,isomaltose and trehalose. The contents ofthose compounds were assayed based onthe comparing peak areas obtained in theexamined samples with those from the re-ference solution (quantitative analysis). To

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Fig. 0. The percent participation of unifloral honey samples used for the study.

make the presentation of the obtained re-sults more comprehensive the followingwas calculated: total sugars, fructose toglucose ratio and total monosaccharides.

The results concerning the contents ofsugars within the groups were processedstatistically by ANOVA. Duncan’s test at asignificance level of α=0.05 was used toexamine the significance of differences be-tween mean sugar contents in the treat-ments under comparison.

RESULTS AND DISCUSSIONIn tables 1-10 and in Fig. 1-10 results

concerning the sugar contents of bee-pro-cessed starch syrup vs. those of honey werecompiled. Statistically significant diffe-rences in the sugar contents to make com-parisons of winter store samples vs. honeysamples were obtained for isomaltose (Ta-ble 7) and for fructose to glucose ratio(F/G) - Table 8, the latter being: 0.82 inwinter store I, 0.97 in winter store II and0.50 in winter store III (nectar honey). Thelowest values of that trait within the honeyswere found for rape honey (0.98 – Fig. 8).The mean isomaltose contents for the re-spective store groups were: 0.48, 0.45 and0.43%. The values were more than twice ashigh as those for the nectar honey andtwice as low as those in honeydew honeysamples. However, because of generallylow isomaltose contents of the products un-der comparison and of the honeys it seemsthat the content of that sugar cannot be agood distinguisher for adulterated honeymade by bees from starch syrups (Table 7and Fig. 7). More significant differenceswere found for sugar contents of bee-pro-cessed starch syrups when they were com-pared with the contents of the same sugarsseparately for nectar and honeydew ho-neys. Significant differences between thecompared treatments – stores vs. nectarhoney – were found for the following traits:fructose content (Table 1), fructose to glu-

cose ratio (F/G – Table 8), and fordisaccharides – sucrose content (Table 3),maltose content (Table 5) and isomaltosecontent (Table 7) and also for totalmonosaccharides (Table 9). Earlier on,Ohe von der and Schönberger (2002),and Liebig (2005) while discussing thecomposition of starch syrups and their suit-ability in beekeeping paid attention to sub-stantial contents of maltose which were re-tained also in bee-processed winter storesdeposited in the combs. The investigatorsalso made note of the fact that the maltosecontent of the stores gradually decreasedwith the concomitant increase in glucosecontent. It was also confirmed by the re-sults of this study. Comparison of the sugarcontent of the stores collected and testedfollowing 2 months (store I) and 7 months(store II) after feeding sugar syrup to beesgave evidence that average maltose contentdecreased from 6.64% to 5.48% (Table 5)and glucose content showed a slight in-crease from 31.05% to 31.59% (Table 2).When samples of starch syrup sampleswere compared with honeydew honey sam-ples the two products differed only for twotraits: greater amounts of the disaccharidesturanose and trehalose were found in ho-neydew honey. The difference was true notonly of the comparison of honeydew honeyvs. stores but also of stores vs. nectar honey(Tables 4 and 6 and Fig. 4 and 6). It is acharacteristic feature of honeydew honeys.

The sugar content values for fructosevaried significantly among the store groupstested. In the samples collected for testingtwo months after feeding (store I) the fruc-tose content averaged 24.89% whereas inthe samples collected 7 months after thesyrup was fed to bees (store II) the averagecontent of that simple sugar was 30.71%.Likewise, the fructose to glucose ratio waslower in stores samples collected 2 monthsafter the feeding (store I) date averaging0.82 (Table 8). However, after 7 months itreached a value characteristic of rape


honey (Table 8 and Fig. 8). With the pas-sage of time, from the date on which thesyrup was fed to bees onwards, the contentof the disaccharide sucrose significantlydeclined from an average of 5.66% to2.30% (Table 3). It is readily evident evennow that store II (centrifuged in the spring,seven months after the bees were fed)meets the requirements as to carbohydratecontents set down by the regulatory docu-ments concerning honey quality currentlyin force. According to those requirementssucrose content must not be higher than 5%

and total saccharides content must not belower than 60% for nectar honeys and 40%for honeydew honeys. The average contentof fructose (30.71% - Table 1) and of glu-cose (31.59% - Table 2) gives a productthat contains 62.3% monosaccharides sothat it also conforms with the standing re-quirement for honey. In Table 9 and inFig. 9 the compiled results for that parame-ter average 71.74% for nectar honeys and65.39% for honeydew honeys. Thus itseems justified to verify those requirements

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Studied materialFructose content %

Mean From To

Starch syrup 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

1) Winter store I - material collected from honeycombs of bee colonies two monthsafter starch syrup was fed to bees in the autumn;

2) Winter store II - material collected from the same bee colonies following seven monthsof feeding of bees (spring of 2005);

3) Winter store III - samples of winter stores which crystallized in honeycombs collected mostof the time in early spring of 2006 (sent in by beekeepers from different regions of Polandfrom the season of 2005/2006);

4) abcd - differences statistically significant between means in rows at α=0.05.

T a b l e 1Fructose content of winter stores made from starch syrup by the bees vs. that of nectar and

honeydew honeys.

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


Studied materialGlucose content %

Mean From To

Starch syrup 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

* For explanation: see Table 1.

T a b l e 2Glucose content of winter stores made from starch syrup by the bees vs. that of nectar and

honeydew honeys.

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

Studied materialSucrose content %

Mean From To

Starch syrup 27.32 25.30 29.30

Winter store I1)* 5.66 c4) 4.31 7.17

Winter store II2) 2.30 b 0.92 3.88


Nectar honey 0.49 a 0.17 0.63

Honeydew honey 1.26 ab 0.85 1.65


T a b l e 3Sucrose content of winter stores made from starch syrup by the bees vs. that of nectar and

honeydew honeys.

30

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

Studied materialTuranose content %

Mean From To


Winter store I1)* 0.98 a4) 0.63 1.44



Nectar honey 1.15 ab 1.02 1.31

Honeydew honey 1.83 c 1.70 2.05


T a b l e 4Turanose content of winter stores made from starch syrup by the bees vs. that of nectar and

honeydew honeys.

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


Studied materialMaltose content %

Mean From To

Starch syrup 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


Honeydew honey 3.61 ab 3.35 3.90


T a b l e 5Maltose content of winter stores made from starch syrup by the bees vs. that of nectar and

honeydew honeys.

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

Studied materialTrehalose content %

Mean From To


Winter store I1)* 0.41 a4) 0.30 0.51

Winter store II2) 0.66 a 0.62 0.72



Honeydew honey 1.71 b 1.25 2.00


T a b l e 6Trehalose content of winter stores made from starch syrup by the bees vs. that of nectar and

honeydew honeys.

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

Studied materialIsomaltose content %

Mean From To


Winter store I1)* 0.48 b4) 0.44 0.51




Honeydew honey 0.88 c 0.83 0.95


T a b l e 7Isomaltose content of winter stores made from starch syrup by the bees vs. that of nectar

and honeydew honeys.

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


Studied materialF/G

Mean From To


Winter store I1)* 0.82 b4) 0.68 1.05

Winter store II2) 0.97 c 0.89 1.05


Nectar honey 1.19 d 1.17 1.21

Honeydew honey 1.18 d 1.16 1.20


T a b l e 8Fructose/Glucose (F/G) ratio of winter stores made from starch syrup by the bees

vs. that of nectar and honeydew honeys.

Studied materialMonosaccharides content %

Mean From To

Starch syrup 25.60 24.70 26.30

Winter store 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


T a b l e 9Monosaccharides content of winter stores made from starch syrup by the bees


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

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

Studied materialSaccharides content %

Mean From To

Starch syrup 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


Nectar honey 76.92 b 75.96 77.49

Honeydew honey 76.03 b 74.05 77.55


T a b l e 1 0Total sugar content of winter stores made from starch syrup by the bees


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

when the domestic regulatory documentsare revised.

The investigations of the spectra of su-gars in syrups and honeys by means of as-says using two chromatograph techniques -GC and LC were also presented by theteam of Cotte et al. (2003) of France.Based on the results obtained they statedthat as small an addition of commercialsyrups to honey as 5 – 10% is already de-tectable. The investigators singled out thefollowing most suitable traits that distin-guish syrups from honey: fructose content,glucose content, fructose to glucose ratio,and also the ratios of sucrose to malotose,maltotoriose to total trisaccharides andmaltotriose to turanose. Based on the re-sults of this study the same traits would bechosen, although without such trisaccha-rides as maltotriose and others because theHPLC method with refractometer detectorand the column selected for this study didnot allow good separation of sugars withinthat range nor did they permit their accu-rate quantitative determination. However,the team of Cotte did not investigate thesugar composition of syrups once theywere processed by bees.

The problem of the crystallization ofstores following their deposition in thecombs by bees as winter food requires sepa-rate treatment. The crystallization occurredonly in a part of apiaries that were fed starchsyrup as winter store in the season of2005/2006. The analysis of the content ofindividual sugars in the samples of thosestores (store III) aided the understanding andthe explanation of the problem. It turned outthat in that group of samples the content offructose was much lower and the content ofglucose was higher. It caused that sugar tocrystallize in the comb cells. According toOhe von der and Schönberger (2000)the critical solution saturation point withglucose is 32 g/100 g and above that con-centration glucose crystallization occurs. Inthe tested samples (store III) the glucoseconcentration was on average as high as38.98% (Table 2).

CONCLUSIONS1. Lower than in honey fructose content

(below 32%), high (over 5%) maltosecontent and a low F/G ratio averaging0.76 vs. 1.18 in honey samples may beused as a distinguisher to identifyproducts processed by bees fromstarch syrups.

2. The store centrifuged in the spring(seven months after the syrup was fedto bees) reached a carbohydrate con-tent in conformance with the require-ments in the valid regulatory docu-ments concerning honey quality. A morein depth analysis of those requirementsseems to be required while revising thecurrent standard PN-88/A-77626 „Miódpszczeli” (Honeybee Honey).

3. In the apiaries in which the crystalliza-tion of the starch syrup-derived storeoccurred the conditions had arisenwhich were more propitious for thehydrolysis of complex sugars mainlymaltose to glucose such as favourableweather conditions, earlier feeding ofbees exceptionally strong bee colonies.Previously, such conditions favouredgood wintering of bee colonies whichwere fed sugar syrup (sucrose solu-tion) but it was not true of this case. Itwas because the bees broke down themaltose-enriched syrup so thouroughlythat it led to a considerable buildup ofglucose and its consequent crystalliza-tion. In those samples glucose concen-tration was found to average 38.98%.

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BADANIA CHROMATOGRAFICZNE (HPLC) SK£ADUCUKRÓW W MIODACH I ZAPASACH NA ZIMÊ

WYTWORZONYCH PRZEZ PSZCZO£Y Z SYROPÓWSKROBIOWYCH

R y b a k - C h m i e l e w s k a H .

S t r e s z c z e n i e

Okreœlono sk³ad cukrów w trzech grupach produktów: w syropie skrobiowym (maltozowym)produkowanym w Polsce ze skrobi pszenicy (który od kilku lat pszczelarze próbuj¹wykorzystywaæ jako pokarm dla pszczó³); w wytworzonym z tego syropu przez pszczo³ypokarmie stanowi¹cym zapas na zimê oraz w miodzie. Zapasy zimowe analizowano w trzechpodgrupach: podgrupa I - materia³ pobrany z plastrów rodzin pszczelich po dwóch miesi¹cachod jesiennego podkarmiania pszczó³ syropem (koniec paŸdziernika 2004); podgrupa II –materia³ pobrany z tych samych rodzin pszczelich po siedmiu miesi¹cach od podkarmiania(wiosn¹ 2005 roku) i podgrupa III - próbki zapasu zimowego, który skrystalizowa³ w plastrach,pobierany najczêœciej wczesn¹ wiosn¹ 2006 (przysy³any przez pszczelarzy z ró¿nych rejonówPolski z sezonu 2005/2006).

Badania zawartoœci cukrów wykonano metod¹ HPLC z detektorem refraktometrycznym wgBogdanova i in. (1997). Oznaczono i porównano zawartoœci nastêpuj¹cych cukrów: glukozy,fruktozy, sacharozy, maltozy, izomaltozy, turanozy i trehalozy.

W porównywanych wynikach zawartoœci poszczególnych cukrów w zapasach utworzonychz syropu i w miodzie odnaleziono istotne ró¿nice. Rutynowe badania sk³adu cukrów metod¹HPLC mog¹ byæ pomocne przy identyfikacji produktów wytworzonych przez pszczo³yz syropów maltozowych. Wyró¿nikiem dla tych produktów by³a ni¿sza w stosunku do mioduzawartoœæ fruktozy (poni¿ej 32%); wysoka (ponad 5%) zawartoœæ maltozy oraz niski stosunekzawartoœci fruktozy do glukozy (F/G), œrednia z badanych podgrup – 0,76, podczas gdyw próbkach miodów wartoœæ ta wynosi³a œrednio 1,18. Najni¿sze wartoœci tego parametru dlamiodów charakteryzowa³y miód rzepakowy i wynosi³y œrednio 0,98.

Zosta³ te¿ wyjaœniony problem krystalizacji zapasów z syropu maltozowego po z³o¿eniu ichprzez pszczo³y w plastrach jako pokarmu na zimê. Krystalizacja wyst¹pi³a tylko w czêœci pasiekkarmionych na zimê 2005/2006 syropem maltozowym. Okaza³o siê, ¿e w tej grupie próbek wstosunku do pozosta³ych (nieskrystalizowanych lub tylko czêœciowo skrystalizowanych)zawartoœæ fruktozy by³a istotnie ni¿sza, natomiast wy¿sza zawartoœæ glukozy. Wg Ohe vonder i Schönbergera (2000) punkt krytyczny nasycenia roztworu glukoz¹ wynosi 32 g/100 g,powy¿ej tego stê¿enia nastêpuje jej krystalizacja. W omawianych próbkach zapasów stê¿enieglukozy wynosi³o œrednio a¿ 38,98%. Spowodowa³o to krystalizacjê tego cukru ju¿w komórkach plastrów. W pasiekach, w których nast¹pi³a krystalizacja zapasu z syropuskrobiowego (maltozowego) zaistnia³y bardziej sprzyjaj¹ce warunki enzymatycznej hydrolizycukrów z³o¿onych do cukrów prostych. Szczególne znaczenie ma tu rozk³ad maltozy, którejw tym syropie by³o stosunkowo du¿o – oko³o 20%. Proces ten powodowa³ szybki wzroststê¿enia glukozy, a losowo bardziej korzystne warunki pogodowe, wczeœniejsze karmieniepszczó³ na zimê, wyj¹tkowo silne rodziny i inne czynniki, przyœpieszaj¹c proces enzymatycznejhydrolizy cukrów z³o¿onych, spowodowa³y szybk¹ krystalizacjê wytworzonych zapasów.Pszczo³y tak dok³adnie przetworzy³y podany im syrop, ¿e doprowadzi³o to do zbyt du¿egostê¿enia glukozy i w konsekwencji do jej krystalizacji w komórkach plastrów.

S³owa kluczowe: miód, syrop skrobiowy, zimowanie pszczó³, wêglowodany, krystalizacja,HPLC, fa³szowanie.

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