7° Encontro Nacional de Cromatografia

Grupo de Cromatografia Sociedade Portuguesa de Qufmica

Departamento de Qufmica e Bioqufmica Faculdade de Ciencias da Universidade do Porto

9 a 11 de]aneiro de 2012

Titulo

7Q Encontro Nacional de Cromatografia

Coordena~ao

Aquiles Araujo Barros Jose Ant6nio Rodrigues Paulo Joaquim Almeida Luis Ferreira Guido M aria Ines Rocha Luis Moreira Gon~alves Manuela Maria Moreira lnes Mar ia Valente Daniel Oliveira Carvalho Jose de Sousa Camara Jose Manuel F. Nogueira

lmpressao e acabamento

Graficos Reunidos Rua Alvares Cabral, 22-32 4050-040 Porto

Edi~ao

Sociedade Portuguesa de Qui mica

Uala

J;111ciro de 20 12

Ti•·agem

275 exemplares

ISBN

978-989-97 66 7-0-9

Deposito legal

338101/11

7° Encontro Naciona l de Cromatografia

in dice

Comprehensive two-dimensional gas chromatography-mass spectrometry (GCxGC-TOFMS}

.111alysis of volatiles in Brazilian Capsicum spp. chili peppers

I I PLC-DAD-MS analysis of Tempranillo grape seeds f rom different geographical origin

Cafc~lol , kahweol and related esters profi le in espresso coffee

On the study of historica l Arraiolos tapestries- a ch romatographic and spect rometric approach

C h.11.1tlcrization of phenolic and polysaccha ridic ext racts of Ganoderma lucidum by

c h1omatographic techniques

/\nlloxldants in Pinus Pinaster and mycorrhizal fungi during the early steps of symbiosis

< lirorna tographic techniques to obtain the biomolecu les prof ile of inedible wild mushrooms

w ith .1nlioxidant value

~l'."ona l variabili t y of essential oi ls of Lavandu/a luisieri

/\n overview of the nutrients and non-nutrients present in the wild mushroom species most

.1 pp1 ('Ciatcd in the Northeast of Portugal

( ,C MS <111alysis of Portuguese propolis

Cylilll~ mulliflorus: phenolic characterization by HPLC-DAD, ESI-MSn and NMR combined

nlt'thods

Ml'lll/10 oquat.ica: source of flavanone glycosides

V.lllcl~llon of a NP-HPLC-FLD method for tocopherol analysis in fresh lettuce

C l hll •lclcdzation of volatile compounds of passion fruit species of semi-arid region in t he

1\1 oll lll,lll Norl hcast

Vol. 1lllc composi tion of Portuguese Oak and Portuguese Chestnut wood chips: their relevance

lo1 <H'nO IOf :l ~d l purposes

0 111 ' cl11111'""0" dl and comprehensive two-dimensional gas chromatography analysis of chestnut

.ul! i o,1k wood chips with different toasting degrees

No·w Ill' 11 1'1 C UV mcthot.l for the quant ification of napthodianthrones from Hypericum species

( h,u ,,< tt•rlt<illon o f Opuntia ficus spp. fractions obtained using Sephadex LH-20.

I if 'h' llllhhllion of bisphcnol A in mussels using dispersive liquid-liquid microextraction and gas

t lll tll lloii OHfoiphy 111 .1~~ ~pcctromc lry

Clpl lmlt.lllon of m \ llu dcrivilliJa tion dispersive liquid-liqu id microextraction for simultaneous

•·xt1 o11 tlo11 ul bh phC'nol 1\ ,tncl b1-.plwnol B in tuna canned fol lowed by gas chromatography-

l io•vc•lnpnlo'llt . \lltl vo~hd ,lllon ol .1 •.lmplt• . 111cl ~cn~i tive t-IPLC/FLD method for N-methyl

1 •ll l foi lll oi lo ". tll ", lif ldo ", ill .l! illlioil ll·,•,llf"• i>.l'•f'<i 011 ci' '• I H 'I ~ ivr <'XIrilrlion by QuEChERS

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7° Encontro Nacional de Cromatografia

indice

New strategy to enhance the extraction efficiency of polyphenols from dietary vegetables using

a modified QuEChERS combined with UHPLC-PDA system

A selective and fast method for the quantitative determination of chloroanisoles in wines using

solid phase microextraction and gas chromatography-quadrupole mass spectrometry

Development of a QuEChERS-based extraction method for Ochratoxin A analysis in bread by LC­

FLD

GC-FID and GC-MS as a tool to screen the influence of wood ageing technologies in the brandies

odourless and odourant compounds

Honey sugars analysis by ion chromatography method with Integrated Pulsed Amperometric

Detection (IPAD)

Ocurrence and bioacessibil ity of nitrates in baby foods

Avalia~ao dos nfveis de contamina~ao com patulina em diferentes fases do processamento

industrial da fruta

Desenvolvimento de um metodo de analise de melamina em produtos alimentares atraves de

M ISPE-HPLC-DAD

Quantification of wine ellagitanins by LC-ESI-MS/MS-mult iple reaction monitoring

Gas-diffusion microextraction: a deeper understanding of the process

Determination of free and tota l diacetyl in wine by HPLC-UV using gas-diffusion microextraction

Determination of pesticide residues in grapes and wine using Quechers and liquid

chromatography - diode-array detection

Recovery and analysis by HPLC-DAD of natural antioxidants from brewer's spent-grain

Analysis of acids and sugars in f ruit-based drinks by SEC-l:JV-RI

Analysis of honey main sugars by size-exclusion chromatography

Monitoring malolact ic fermentation using HPLC-UV-RI

Determination of chloropropanols in water and food samples using ult rasound-assisted

dispersive liquid-liquid microext raction with derivatization and GC-MS detect ion

Arsenic speciation in rice and fish using HPLC-ICP-MS

Solid-phase extraction fol lowed by dispersive l iquid-liquid microext raction for the sensitive

determinat ion of ethylphenols species in w ine

Fast detect ion of ~-lactam antibiotics in mi lk by isocratic HPLC elution

Chromatographic analysis of macro and micronutrients in the most widely appreciated

cultivated mushrooms

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P86. Analysis of honey main sugars by size-exclusion chromatography

Mara E. B. C. Sousa0·*, Ant6nio M. Peresa.b, Lufs G. Diasa, forge Sa Morais0, Letfcia Estevinho0 , Adelio A.S.C. Mach a doe

aCIMO- Escola Superior Agraria, lnstituto Politecnico de Bragan~a. Portugal, b LSRE - Laboratory of Separation and Reaction Engineering- Associate Laboratory LSRE/LCM, Escola Superior Agraria, lnstituto

Politecnico de Bragan~a. Portugal, cLAQUIPAI - Departamento Quimica, Faculdade de Ciencias, Universidade do Porto, Portugal

•mcbdias@iol.pl

The main constituents of honey are sugars, mainly the monosaccharides fructose and glucose (reducing sugars), which account for 80% of the total mass, while the disaccharides (especially sucrose and maltose) represent 10%. In general, fructose predominates (giving a sweeter taste to honey than sucrose), although, in some honeys, glucose (that assigns a less sweet taste than sucrose but more than maltose) may be the main sugar. The fructose/glucose mass ratio is a parameter that allows the identification of possible adulterations of honey, by signaling abnormal levels of glucose due to the addition of commercial glucose or syrups. lt also allows assessing the crystallization trend of the honey, although the glucose/water ratio provides a more accurate indication of this behavior. Therefore, for marketing purposes, the quantification of glucose and fructose in honey is a key factor for establishing its quality. This work reports the validation of a method for determination of honey sugars by SEC to be used as control in the study of their determination with an electronic tongue. A set of honey sa mples was provided by the Portuguese National Federation of Beekeepers to obtain a national sampling which showed the natural composition variability of Portuguese honeys. The analysis were carried out with HPLC equipment from Varian and a Supelcogel C-610H column, selected because the elution does not require organic solvents. The target compounds to analyze were glucose and fructose for which well separated peaks were found in the chromatograms, which included other two, for a mix of polyssacharides and unknown substance(s). The results of a thorough validation showed that the method can be employed for quantification of these two sugars in honey samples.

156

ANALYSIS OF HONEY MAIN SUGARS BY SIZE-EXCLUSION CHROMATOGRAPHY Mara E. B. C. Sousa&.", Ant6nio M. Pere~·~'~, Lais G. Dias9. Jorge SB Morais&,

Leticia Eslevinhoflr Adf:lio A.S.C. Machadac

"CIMO • Esooaa ~dor Agnirfa. tnstltuto Politecn.co de Bragan98-, Portugal. • LSRE • Laboratory of Separallon and Reaction Engineering • As9oclale Laboratory LSREII..CM, Escota Superior Aglliria, tns.tituto Pollte<:nlco de Bragan~ . Portugal, ~LAOUIPAI • Oepartamento Ootmlca. f aC\Ad&de de CiAnclas. llri'Versld&de do Potto. Portugal

Introduction Main constitu&nts of honey are sugars:

' 80% fructose and glucose: • 10% d~acchandes (especially sucrose and maltose).

Fructos& pr&dominatGs sn honey In some. glucose may be tOO main sugar.

Fructos&/glucos& mass tabO:

ldentJiicabOn of possl.ble adultoratioos of honey, • >1 .3 will crystallize stowty or remam liquid.

Objectives HPLC method vahdabon for honey sugars determination by SEC {slz&-exctusion Chromatography)

Referanoo method Jn the study of their detetmination with an atactronlc tongue.

Samples !lar.eys l'lfl)•.'ld!ld by rMP • Pvtt;'}uesu c Cd111<tt :;(1 uf Bcu~:i

P.u~t.':J.li fr<1~1 dtllllfCfll P .t.~3 11:91tlb

Sa!"'!p!llS lltC1)3f3!11Yl-DILUTION.

HPLC equipment 'Ja_"lan Pro!ilar 22' .. 1 Pump Va."lar Rl-·1 lrefrat:we 1ndcx OOtcctori

•metx:t!as@lol.pt

Results 1. COO"'pound id.entification - ret&ntlon timo-. 2. Calibration- glucose and fructose. 3. Study of ACCURACY ao>d PRECISION:

• 2 quality conttol solutloi\S; • 3 honey sampl&s (different cotom).

4. T&n different honeys aJlalysed.

- , - · ~-1 -· ' •

- · • •

--­·---:.,Q,Ii!)osaothandes ?

Orill - ll"iiiiii. (Q"CJCC11t'l«<

cm...tr~o»tTJ.II(mti.Y'tf1)M

Ot·~tcl#;lll,) Qt.q.mtlmtt~lru

---Olsacch:afldas - fnsacttla1h~ ..-1

RhCO:ly'le nz: .• Manual ;t!JI)CIOf ''l'llloop (11 LU IJl Softwa.-o Sic\' ciY•)matograp~y I'IC'b .. .:al1!:4. ,•crMon 6.4 Jvtes.. lt:.a 1 thrt.tr..atograph)• culuz"ln o,•cl'l

(lu:My tontml SOiuliom fo!e:!~W't Gwmrti .stnUit ... M ltllrt ...

~~bl~

-""·" ...... .... COLUMN Supelcogel C 610H d•3Ccrr x 7.amrr 01

GREEN CHEMICAL ANALYSIS

HPLC conditions C~h.tr!l !CMPQ•rlluu::. JCflC £lull!:' cl.lt'll)t6tl10: l'lf"oi phospllor!c :.cid aqultO'JS St>lllilon Eiuur' c t;hOil •socf!IJc: Flux; UA ll'h .. •'l~ Eluonuemil!IUIJ1e. 40•c

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ot iil •• l•ll

,, iiUi

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i!.\6 di t((

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""""' hrenntrtll fuiU~>~ Aiiii~'t IQJM Gfl!lrif'('ffli &in:GO IQn., ~UfOtl .. ._. """"""' WO:~~' ~»<Oln•Kt 'btc!a•f "~ "'""' ........ ((n(l!f~Jdt'l\ ~dt'II!JI '"""' " :l\ i' 4Mllttlfslt:ol ,..,., 9tlto:llfcnoo; ~·~~~~~!! dcr.u~·~~

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i£32 ......... QIS ijiJ Otil I) Olll V•.l.- 36 dS ib f fll.tiH "' '" '" " " h llt;W '" •• •• ..,_

"" .. " O:~•<!Uilfti.Uf.Uidii~U ,,..,., ,., .. .,

Conclusions

. -----~

·~.---------------,

I~

1: •

The CNilrall calibrallons are acceplable ~ Re$ults or accuracy and precls!oo are atceplable . • TOO meltlod can be Mlployed t« quaollflcallon of these two sugn io hOOOf samplE!$. Fructosa 1n ho08)' a~1,qys great et than glucose ~ f ruclosetglucose mass ra11o in honey' highflr than 1.34 . • HOIWfS will ~amze slatdy ot remain llqUJ-11.