Journal of Chromatography A, 1208 (2008) 1624

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Journal of Chromatography A

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Applica extdeterm geliquid c ss s

P. Plaza B J. L.Department of

a r t i c l

Article history:Received 30 JuReceived in reAccepted 18 AAvailable onlin

Keywords:PesticidesWineBeerLiquid phaseHollow breUHPLCMS/

opedxtrac

spectple t

ograption such as type of organic solvent, extraction time and agitation rate have been optimized. The extractionmethod has been validated for several types of alcoholic beverages such as wine and beer, and no matrixeffect was observed. The technique requires minimal sample handling and solvent consumption. Using

1. Introd

Nowadcides are umolds, ancides are uagriculturcessed anCommunifor viniferfor pesticisetting spThereforeof sampleof these panalyticalto monito

Basicalmatograp

CorrespE-mail a

0021-9673/$doi:10.1016/microextraction

MS

optimum conditions, low detection limits (0.015.61g L1) and good linearity (R2 > 0.95) were obtained.Repeatability and interday precision ranged from 3.0 to 16.8% and from 5.9 to 21.2%, respectively. Finallythe optimized method was applied to real samples and carbaryl, triadimenol, spyroxamine, epoxicona-zole, triumizol and fenazaquin were detected in some of the analyzed samples. The obtained resultsindicated that the new method can be successfully applied for extraction and determination of pesticidesin alcoholic beverages, increasing sample throughput.

2008 Elsevier B.V. All rights reserved.

uction

ays, a wide range of herbicides, insecticides and fungi-sed in grape production to protect against insects, fungi,

d other agents that may affect crop yield [1]. When pesti-sed in or onplant products, residues can occur on the rawal commodities and they can be transferred to the pro-d nished food products [2]. Despite of several Europeanty Directives have xed maximum residue limits (MRLs)ous grapes [3], no uniform limits have been establisheddes in wine, although there is a worldwide trend towardsecic MRLs in wine, ranging from 0.01 to 2mgL1 [4]., thedeterminationofpesticides residue levels in this types is of special concern to ensure the safety consumptionroducts. In this sense, the development of multiresiduemethods for the determination of pesticides is essentialr and control the use of agrochemical in vineyards.ly capillary gas chromatography (GC) [5,6] or liquid chro-hy (LC) [7,8] have been the selected techniques for the

onding author. Tel.: +34 950015823; fax: +34 950015483.ddress: rromero@ual.es (R. Romero-Gonzlez).

analysis of pesticide residues in beverages, although capillary elec-trophoresis (CE) has been also used [9]. However, in the last fewyears, the use of LC coupled with mass spectrometry detectors(LCMS), such as single quadrupole, triple quadrupole or timeof ight [10] has improved the sensitivity and has been rapidlybecoming an accepted technique in pesticide residue analysis forregulatory purposes. Nevertheless, one of the main problems asso-ciated with this combination is the ion suppression due to matrixeffects, so the selected extraction technique should minimize thiseffect [11].

Traditionally, routine methods involve several sample prepara-tion steps such as extraction, clean-up and concentration beforeinstrumental analysis, using liquidliquid extraction (LLE) or solidphase extraction (SPE) as extraction techniques [6,9,12]. However,these sample preparation steps are tedious, time-consuming andthey consume large amount of solvents. On the contrary, newmicroextraction techniques such as solid phase microextraction(SPME) [13,14], stir bar sorptive extraction (SBSE) [15,16] and liq-uid phase microextraction (LPME) [17,18] have been introducedbecause they are easy, fast and solvent-free or less organic solventconsumption techniques. Among these techniques, liquid phasemicroextraction using hollow bre membranes (HF-LPME) [19]provides mechanical stability and protection to the organic solvent

see front matter 2008 Elsevier B.V. All rights reserved.j.chroma.2008.08.059tion of hollow bre liquid phase microination of pesticides in alcoholic beverahromatography coupled to tandem ma

olanos, R. Romero-Gonzlez , A. Garrido Frenich,Analytical Chemistry, University of Almeria, E-04071 Almeria, Spain

e i n f o

ne 2008vised form 14 August 2008ugust 2008e 20 August 2008

a b s t r a c t

An alternative method has been develusing hollow bre liquid phase microematography coupled to tandem massPesticides were extracted from the samdesorbed inmethanolprior to chromat/ locate /chroma

raction for the multiresidues by ultra-high pressurepectrometry

Martnez Vidal

to determine more than 50 pesticides in alcoholic beveragestion (HF-LPME) followed by ultra-high pressure liquid chro-rometry (UHPLCMS/MS), without any further clean-up step.o the organic solvent immobilized in the bre and they werehic analysis. Experimental parameters related tomicroextrac-

P.P. Bolanos et al. / J. Chromatogr. A 1208 (2008) 1624 17

Fig. 1. Schema sampin methanol; (

because theeffective, lovides an exextracts. Onthat the accwithin the presence ofsome selectmolecules,extracted.

Basicallyimpregnateextracted bbic organicthe other hsolvent immaqueous phWhereas twcompoundsbeen preferCE as analymode, the oand the lumwork, whichof the holloadsorbed inGC analysisthe three phpatible solvthe acceptodesorption

The aimsimple extrof 51 pesticiultra-high pto tandemmatrix effection techniq

The comas HF-LPME

e ind for

erim

agen

tiedGm

ver,lways (wrepationundeolutied bandtic of HF-LPME extraction: (a) immobilization of the organic phase in the bre; (b)e) removing the bre before chromatographic separation.

use of a membrane or hollow bre, which is simple,w cost, uses microliters of organic solvents and pro-cellent sample clean-up ability, obtaining very cleane of the main advantages of this technique over SPME iseptor solution in hollow bre was effectively protectedbre which served as lter, while SPME suffers from thesample particles [20]. Besides, HF-LPME can also showivity because of the pores in its wall. In this sense, largewhich can be soluble in extracting solvent, may not be

, the piece of porous polypropylene hollow bre isd with a water-immiscible solvent and the analytes arey passive diffusion from the sample into the hydropho-solvent supported by thebre (twophaseHF-LPME). Onand, the analytes can be extracted through the organicobilized in the pores of the bre and further into a newase in the lumen of the bre (three phase HF-LPME).o phase mode has been mainly used for hydrophobic

increasmetho

2. Exp

2.1. Re

CerstorferHannowere apoundwere pdissolustoreddard spreparMeOH, which were analyzed by GC, three phase mode hasably used for ionisable compounds [21], using LC ortical techniques [22]. Traditionally, in the two phaserganic solvent comprises the pores of the membraneen inside the hollow bre, although there is a recentonly use the organic solvent immobilized in the pores

w bre as acceptor phase [23], and the organic solventthe bre was desorbed after the extraction step prior

. This procedure could be an interesting alternative toase system when the bre can be desorbed in a com-ent for LC analysis, minimizing bre handling due tor is not injected into the lumen of the bre, and only astep is necessary prior LC analysis.of the present study is the development of a new andaction procedure, based on HF-LPME for the extractiondes fromalcoholic beverages, prior to determination byerformance liquid chromatography (UHPLC) coupled

mass spectrometry (MS/MS), possibly eliminating thets normally founded by LCMS/MS when other extrac-ues are employed.bination of a simple and fast extraction technique suchtogether with the use of UHPLCMS/MS permits the

grade solveAcetonitrileBaker (Devanalysis grgrade) werSigma. OtheAldrich, SteBuchs, SwitQ3/2 Accur(200m wobtained frsyringe pluland). Highwas used tsamples.

2.2. Instrum

ChromatUPLCTM sywere achie(100mmle extraction; (c) removing the bre from the sample; (d) desorption

sample throughput and the suitability of the proposedroutine analysis.

ental

ts and materials

pesticide standards were purchased from Dr. Ehren-bH (Augsburg, Germany), Riedel-de-Han (Seelze-Germany) and Chemservice (Milan, Italy). Puritiess >95.0%. Stock standard solutions of individual com-ith concentrations ranging from 200 to 300mgL1)red by exact weighing of the powder or liquid andin 50mL of methanol (MeOH), which were thenr refrigeration (T5 C). Amultipesticideworking stan-on (2g L1 concentration of each compound) wasy appropriate dilution of the stock solutions withstored under refrigeration (T5 C). MeOH (HPLCnt) was obtained from Sigma (St. Louis, MO, USA).(ACN, HPLC grade solvent) was supplied by J.T.

enter, The Netherlands). Ethyl acetate (EtAc, residueade) and dichloromethane (DCM, residue analysise supplied by Riedel-de-Han and 1-octanol fromr reagents were trioctylphosphine oxide (TOPO; 99%;inheim, Germany), dihexyl ether (DHE; 97%; Fluka,zerland) and NaCl (99.5%; Panreac, Barcelona, Spain).el PP hydrophobic polypropylene hollow bre tubingall thickness, 600m i.d., and 0.2m pore size) wasom Membrana GmbH (Wuppertal, Germany). 10-Lngers were provided by Hamilton (Bonaduz, Switzer-ly puried water (milli-Q Millipore, Bedford, USA)hroughout for the preparation of mobile phase and

ents and apparatus

ographic analyses were performed with an Acquitystem (Waters, Milford, MS, USA) and separationsved using an Acquity UPLCTM BEH C18 column2.1mm, 1.7-m particle size) from Waters.

18 P.P. Bolanos et al. / J. Chromatogr. A 1208 (2008) 1624

Table 1Logarithm of octanolwater partition coefcient (logKow), retention time windows and MS/MS conditions

Pesticide logKow RTW (min) Cone voltage (V) Quantication transitiona Conrmation transitiona

SimazineDesethyl terbuCarbarylMonolinuronChlorotoluronMetobromuroAtrazineMetazachlorLenacilFensulfothionIsoproturonDiuronAzaconazoleIodosulfuron mBensulfuron mDiethofencarbSebuthylazinePropazineLinuronSpiroxamineMethiocarbTerbuthylazinePaclobutrazolFlutalonilPromecarbPrometrynPropyzamideTriazophosTepraloxidimIprovalicarbTriadimenolFenhexamideEpoxiconazoleTebutamMetolachlorFenbuconazolDiubenzuronCyprodinilThiazopyrFurmecycloxSpinosadBitertanolPencycuronTrioxystrobinTriumizoleClethodimCycloxydimFluazifop buthSethoxydimHexythiazoxFenazaquin

a Collision e

MS/MSdem quadruinstrumentitive mode.3.5 kV, extrtion temperow 600Lhciation waspressure ofmonitoringvoltages apperformed(Waters). Amany), a Unbath from Jextraction.2.1 3.323.43 35thylazine 1.9 3.533.67 20

1.8 3.543.68 202.2 3.633.77 282.5 3.793.94 25

n 2.4 3.833.96 282.5 3.884.03 302.1 3.894.06 152.3 3.924.10 202.2 3.944.11 302.5 3.974.13 302.8 4.054.20 302.2 4.084.23 20

ethyl 1.1 4.084.28 25ethyl 0.6 4.184.30 40

3.0 4.304.45 203.2 4.334.51 253.0 4.364.53 25

3.0 4.374.54 302.8 4.384.62 253.1 4.444.59 223.2 4.464.60 283.2 4.514.67 253.7 4.524.66 273.2 4.534.68 233.1 4.544.69 203.2 4.614.72 253.3 4.664.80 351.5 4.694.85 203.2 4.704.82 203.2 4.734.86 253.5 4.784.90 253.4 4.824.95 253.0 4.884.98 272.9 4.915.02 203.2 4.915.02 303.9 4.975.07 254.0 4.975.09 203.9 5.055.16 353.6 5.135.26 204.5 5.145.47 304.0 5.295.39 254.7 5.405.45 404.5 5.435.51 305.1 5.515.59 235.1 5.535.62 201.4 5.535.62 22

yl 4.5 5.605.67 354.5 5.685.76 232.5 5.875.95 255.5 6.266.37 30

nergy (eV) is given in parenthesis.

detection was performed using an Acquity TQD tan-pole mass spectrometer (Waters, Manchester, UK). Thewas operated using an electrospray (ESI) source in pos-The ESI source was set as follows: capillary voltage

actor voltage 5V, source temperature 110 C, desolva-ature 350 C, cone gas ow80Lh1 and desolvation gas1 (both gases were nitrogen). Collision-induced disso-performed using argon (99.999%) as collision gas at a4103 mbar in the collision cell. Themultiple reaction(MRM) transitions and the cone and collision energyplied are summarized in Table 1. Data acquisition wasusing MassLynx 4.0 software with QuanLynx softwareReax-2 rotary agitator from Heidolph (Schwabach, Ger-itronic 320 OP longitudinal agitator and an ultrasoundP Selecta (Barcelona, Spain) were available for sampleAn analytical balance was used in standard preparation.

2.3. Sample

TheHF-Lhollow brinserted intwith 1-octacontainingdure, the viagitator forand plungewas put intvial was plaat approximanalytes frofrom the viaUHPLCMS202.1 >132.1 (18) 202.1 >96.1 (25)202.3 >146.1 (16) 202.3 >78.9 (25)202.1 >145.1 (10) 202.3 >127.0 (30)215.1 >126.1 (18) 215.1 >148.1 (15)213.2 >72.1 (15) 213.2 >46.2 (15)259.1 >170.0 (20) 259.1 >148.1 (20)216.1 >174.1 (18) 216.1 >96.1 (25)278.3 >134.0 (20) 278.3 >210.2 (10)235.2 >153.0 (15) 235.2 >136.0 (30)309.1 >281.2 (15) 309.1 >157.0 (26)207.2 >72.1 (30) 207.2 >165.2 (15)233.1 >72.0 (25) 233.1 >160.0 (18)300.1 >159.0 (23) 300.1 >231.0 (16)530.4 >163.2 (16) 530.4 >390.2 (16)411.3 >149.2 (18) 411.3 >182.2 (18)268.3 >226.3 (10) 268.3 >152.1 (20)230.4 >174.1 (18) 230.4 >95.9 (25)230.4 >188.2 (16) 230.4 >146.6 (20)

249.1 >160.0 (18) 249.1 >182.1 (18)298.3 >144.2 (20) 298.3 >100.1 (33)226.1 >169.2 (10) 226.1 >121.1 (18)230.2 >174.1 (15) 230.2 >96.1 (25)294.2 >70.0 (25) 294.2 >125.0 (25)324.4 >242.3 (25) 324.4 >262.3 (20)208.2 >151.2 (9) 208.2 >109.1 (15)242.0 >157.9 (17) 242.0 >200.3 (25)256.2 >190.0 (16) 256.2 >173.0 (23)314.2 >162.1 (18) 314.2 >119.1 (34)342.4 >250.3 (12) 342.4 >166.1 (20)321.4 >119.1 (15) 321.4 >203.3 (8)296.2 >70.0 (12) 296.2 >99.1 (12)302.2 >97.1 (25) 302.2 >55.1 (30)330.2 >121.1 (20) 330.2 >141.1 (20)234.3 >90.9 (18) 234.3 >192.2 (18)284.3 >252.4 (15) 284.3 >176.3 (23)337.3 >70.1 (25) 337.3 >125.0 (20)311.1 >158.1 (12) 311.1 >141.1 (22)226.0 >108.1 (20) 226.0 >93.1 (20)397.2 >377.2 (28) 397.2 >335.1 (22)252.5 >170.2 (13) 252.5 >110.0 (20)732.7 >142.2 (30) 544.5 >142.2 (25)338.3 >269.3 (10) 338.3 >99.2 (15)329.3 >125.1 (19) 329.3 >218.3 (14)409.3 >186.2 (18) 409.3 >206.2 (14)346.2 >73.1 (17) 346.2 >278.2 (11)360.3 >164.1 (20) 360.3 >268.3 (12)326.3 >280.4 (13) 326.3 >180.2 (20)384.3 >282.3 (20) 384.3 >91.0 (30)328.5 >178.1 (18) 328.5 >282.3 (11)353.3 >228.1 (15) 353.3 >168.1 (20)307.3 >161.2 (16) 307.3 >147.1 (16)

preparation by HF-LPME

PMEprocedure shown inFig. 1waspreparedas follows:es were cut into 2-cm pieces which were completelyo a 10-L syringe plunger. The bre was impregnatednol (1min) and then placed into a 15-mL screw top vial15mL of homogenised sample. In the optimized proce-al was carefully closed and put into a rack in the rotary45min at 90 rpm. After the extraction stage,wine tracesr were removed by using tweezers and only the breo a 2-mL vial containing 1.5mL of MeOH. This secondced into a rack and then in the rotary agitator for 5minately 30 rpm in order to perform the desorption of them thebre to the solvent. Finally, the brewas removedlwith tweezers and 5L of samplewere injected in the/MS system.

P.P. Bolanos et al. / J. Chromatogr. A 1208 (2008) 1624 19

2.4. UHPLCMS/MS analysis

Chromatographic analyses were carried out using gradient elu-tionwith amformic acideluent B, w1min). Thisre-equilibraning time oand the colsample extr

2.5. Sample

A varietyfrom localobtained frlyzed followshowing thsamples in

3. Results

Severalphase or twa new one,immobilizehandling anbre is tranlytes are dewas used asing in mindstudy (see Ttwo or thresolvent, diffstripping soof the selec

On the oa previously

3.1. Optimiz

DifferencidesbyHF-The optimizwith 25gdesorptionimmersed i

First, thebrane wasanalytes frovent). The sof the memshould be iso DHE, andwere also tof pesticideresults for mumizole arsolvent, betconsequencbre, althopropyzamidfenazaquin,

ffectlazineked we: 30min (metha

obseere uelatioeringed simnol wol as

t.ringcycl

methin ortratiemes thas n

wellractiitionto 2antlyprope ext, so ful.impthe sg, altagitathe owas cydim (similar results were obtained for the rest of assayeddes), observing that better results were obtained when thead shaker (rotary agitation) was used.n, shaking speedwas studied, and Fig. 3b shows the obtainedfor promecarb and trioxystrobin (the same prole was

ed for the rest of pesticides). It can be observed that extrac-creased with the agitation rate of the sample up to 90 rpm.speed was not selected taking into account the formation ofbles in the hollow bre and the loss of the organic solvent.ring in mind that analytes need time for their mass transferh the sample to the liquid membrane, extraction time is anant parameter that should be optimized. Fig. 4 shows theed results for desethyl terbuthylazine and azaconazol (timefor simazine, and ofurace are similar), diethofencarb andnol (time prole for the rest of pesticides is similar) whenion time was studied between 15 and 240min. It can beobile phase prepared fromMeOH (eluent A) and 0.01%in water (eluent B). The analysis started with 25% ofhich was linearly increased up to 90% in 5min (holdcomposition was held for further 2min. Afterwards, ation time of 1.5min was included and so, a total run-f 11min was obtained. The ow rate was 0.35mLmin1

umn temperature was set at 35 C. Aliquots of 5L ofact were injected.

s

of commercial wine and beer brands were purchasedsupermarkets in Almeria (Spain); other samples wereom home-made productions. All samples were ana-ing the procedure described below and those samples

e absence of the target compounds were used as blankthe preparation of standards and recovery studies.

and discussions

set-ups can be used in HF-LPME, which implies threeo phase systems. Recently, Zorita et al. [23] describedwhich only uses as acceptor phase the organic solventd in the pores of the hollow bre, minimizing sampled reducing extraction time. After the extraction, the

sferred to a vial containing another solvent and the ana-sorbed from the bre to this last solvent. This procedurean alternative to conventional two phase system. Bear-the different polarity of the pesticides selected in thisable 1), this set up could be an interesting alternative toe phase system, because depending on the desorptionerent pesticides could be desorped from the bre, to alvent, so this approach was selected for the extractionted pesticides.ther hand, the chromatographic analysis was based onreported UHPLCMS/MS methodology [24].

ation of the HF-LPME parameters

t parameters that inuence the extraction of the pesti-LPMEwereoptimized, selectingpeakareas as response.ation was carried out using blank white wine spikedL1 of each pesticide, using 2 cm of bre and 1.5mL ofsolvent; this volume enables the bre to be completelyn the solvent (see Fig. 1).organic phase immobilized in the pores of the mem-

studied, as well as the solvent used to reextract them the membrane after the extraction (stripping sol-election of the organic phase immobilized in the poresbrane was selected taking into account that solvent

mmiscible with the sample and stable (low volatility),1-octanol were studied. Furthermore, DCM and EtAc

ested because they are mainly used for the extractions in conventional liquidliquid extraction. The obtainedetazachlor, linuron, terbuthylazine, cyprodinil and tri-e shown in Fig. 2. When 1-octanol was used as organicter results were obtained for most of the pesticides. Ine, it was used as organic solvent immobilized in theugh for some pesticides such as linuron, promecarb,e, pencycuron, triuoxystrobin, uazifop buthyl andbetter results were obtained when DHE was used. It

Fig. 2. Eterbuthywine spition timtime: 10dichloro

can beEtAc w

In rconsidprovidmetha1-octansolven

Beasuch asfuronaddedconcenimprovwhereaTOPO w

It isthe extof addfrom 0signicine, fenthat thofNaClof NaC

Onetion ofshakinrotaryshowstationsethoxpesticioverhe

Theresultsobservtion inHigherair bub

Beathrougimportobtainprolebitertaextractof the organic solvent on the peak area of metazachlor, linuron,, cyprodinil and triumizole. Extraction conditions: 15mL of whiteith 25g L1 of each pesticide; stripping solvent: methanol; extrac-min.; type of agitation: shaking (80oscillationsmin1); desorptionshaking). Abbreviations: DHE: dihexylether; OCT: 1-octanol; DCM:ne; EtAc: ethyl acetate.

rved that lower results were obtained when DCM andsed, maybe due to the volatility of these solvents.n to the stripping solvent, MeOH and ACN were usedthat they are LCMScompatible solvents. Both solventsilar results, but better peak shape was obtained when

as used, so further experiments were carried out usingsolvent immobilized in thebreandMeOHasstripping

inmind that someof the compounds are relativelypolar,oxidim, tepraloxidim, bensulfuron methyl and iodosul-yl (logKow

20 P.P. Bolanos et al. / J. Chromatogr. A 1208 (2008) 1624

Fig. 3. Effect othe peak areaconditions: 15solvent: 1-octtion time: 10mpromecarb (spiked with 2vent: methano(shaking).

observed that short extfor diethofe60min, anddecreased. Tdecreased oto the samping sensitivselected as

Desorptimethanol aand rotarywhen rotartime was evresults for mresultswere

Fig. 4. Extracdiethofencarbspiked with 2vent: methano

Fig. 5. Study oExtraction conorganic solvenagitation spee

obtained wand the sign

o ince. Th

nrmhe onal wsorpextrto

f exted (Tas a

des sand%,Hpestwhery in tf the shaking on the extraction. (a) inuence of the type of agitation onof carbaryl, paclobutrazol, epoxiconazol and sethoxydim. ExtractionmL of white wine spiked with 25g L1 of each pesticide; organicanol; stripping solvent: methanol; extraction time: 30min; desorp-in (shaking). (b) Inuence of the agitation speed on the extraction of

) and trioxystrobin (). Extraction conditions: 15mL of white wine5g L1 of each pesticide; organic solvent: 1-octanol; stripping sol-l; extraction time: 30min; agitation: rotary; desorption time: 10min

at for desethyl terbuthylazine peak area kept constantraction time (1530min) and then decreased; whereas

order ttion timwas cotions. Tthe sigrst de

Theappliedvalue oobtainbility wpesticidiuronthan10tion ofFig. 6,directlncarb and related compounds, peak area increaseduntilwhen extraction time was increased, the signal slightlyhisbehaviourwasdescribedpreviously [27,28], and thef peak area can be due to pesticideswere extracted backle together with the loss of organic solvent. Consider-ity and analysis time, an extraction time of 45min wasoptimal condition.on conditions were also evaluated, using 1.5mL ofs stripping solvent. First, sonication, horizontal shakingagitation were studied. The best results were obtainedy agitation was applied at 30 rpm. Second, desorptionaluated from 5 to 60min, shown in Fig. 5 the obtainedonolinuron, paclobutrazol and fenazaquin (the sameobtained for the restofpesticides). Thesamesignalwas

tion time prole of desethyl terbuthylamine (), azaconazol (),() and bitertanol (). Extraction conditions: 15mL of white wine

5g L1 of each pesticide; organic solvent: 1-octanol; stripping sol-l; agitation speed: 90 rpm (rotary); desorption time: 10min (rotary).

by the optimmination (Fwhen winewhereas thonly the peon the chronique of the

3.2. Study o

Ethanolit is well knin alcoholicthe sampletent. In thisextraction v2.5 to 15mconcentratiextracted amvolumewasresponses win order tono dilution

The useinuence omination ostudied. In talcoholic anmatrix effecated matrf the desorption time formonolinuron, paclobutrazol and fenazaquin.ditions: 15mL of white wine spiked with 25g L1 of each pesticide;t: 1-octanol; stripping solvent: methanol; extraction time: 45min;d: 90 rpm (rotary).

hen short desorption times (510min) were applied,al does not decrease with longer desorption time, so inrease sample throughput, 5minwas selected as desorp-eefciencyofbackextractionof the selectedpesticidesed by performing two consecutive methanol desorp-

btained results indicate that in the second desorption,as always lower than 5% of the signal obtained for thetion, so a second desorption was not necessary.action procedure with the optimized conditions wasspiked blank white wine (25g L1), and the meanraction efciency and relative standard deviation wereable 2), where it can be observed that good repeata-chieved. It must be indicated that although for someuchas simazine, desethyl terbuthylazine, chlorotoluron,bensulfuron methyl, extraction efciency was lower

F-LPME is anefcient clean-upprocedure for the extrac-icides from alcoholic beverages as it can be observed ine a spiked blank white wine (25g L1) was injectedhe UPLCMS/MS chromatogram (Fig. 6a) and extractedizedHF-LPMEprocedure prior chromatographic deter-

ig. 6b). It can benoted that pesticideswere not observedwas directly injected due to the matrix components,

e extracted wine provide a clean chromatogram, wheresticides were detected, and no interfering peaks appearmatogram, indicating the efciency as clean-up tech-HF-LPME.

f matrix effect on the extraction process

is one of the major constituents of wine and beer andown that it can inuence the extraction of pesticidesbeverages [29], and sometimes it is necessary to diluteprior to extraction in order to reduce alcoholic con-work, the effect of dilution was studied, xing a totalolume of 15mL, adding several volumes of wine (from

L) and lled with water up to 15mL, using a nalon of pesticides of 25g L1 in the nal solution. Theounts were approximately the same when the sample

between5and15mL (datanot shown), obtaining lowerhen only 2.5mL, as it was observed previously [13], so

minimize sample handling and to increase sensitivity,of sample was carried out.of HF-LPME could be affected by matrix affect, so thisver the extraction process and UHPLCMS/MS deter-f the pesticides in different alcoholic beverages, washiswork, ve beverages (white, red and sparklingwine,d non-alcoholic beer) were selected for evaluation ofct, analyzing the extraction efciency when the indi-ices were spiked with 25g L1 of pesticides. Table 2

P.P. Bolanos et al. / J. Chromatogr. A 1208 (2008) 1624 21

Table 2Extraction efciency and mean relative recoveries of pesticides in different matrices spiked at 25g L1

Pesticide %Ea Relative recovery (%)b

SimazineDesethyl terbuCarbarylMonolinuronChlorotoluronMetobromuroAtrazineMetazachlorLenacilFensulfothionIsoproturonDiuronAzaconazoleIodosulfuron mBensulfuron mDiethofencarbSebuthylazinePropazineLinuronSpiroxamineMethiocarbTerbuthylazinePaclobutrazolFlutalonilPromecarbPrometrynPropyzamideTriazophosTepraloxidimIprovalicarbTriadimenolFenhexamideEpoxiconazoleTebutamMetolachlorFenbuconazolDiubenzuronCyprodinilThiazopyrFurmecycloxSpinosadBitertanolPencycuronTrioxystrobinTriumizoleClethodimCycloxydimFluazifop buthSethoxydimHexythiazoxFenazaquin

a Mean extrb Relative st

shows the rused as repthe differenmatricesdodure. Furthethe reliabiliThis meansmatricesevwithout ma

3.3. Validat

The optiearity, accuWhite wine Red Wine

3.6 (6.4) 83.3 (7.2) 86.1 (14.7)thylazine 2.5 (3.2) 99.2 (5.7) 96.8 (14.1)

4.9 (3.6) 73.5 (10.8) 76.3 (5.9)4.3 (3.6) 97.7 (12.2) 75.8 (13.3)3.6 (8.2) 97.2 (8.4) 103.9 (4.3)

n 4.9 (5.3) 94.3 (6.4) 87.3 (15.6)7.0 (10.5) 94.0 (7.8) 97.4 (9.8)2.2 (10.7) 80.0 (3.8) 87.3 (7.8)3.4 (6.3) 73.5 (5.7) 108.2 (5.6)6.9 (12.6) 97.4 (8.6) 97.1 (6.3)4.2 (13.8) 87.5 (10.5) 102.5 (5.3)4.5 (17.0) 102.8 (9.9) 106.0 (9.5)2.6 (16.0) 85.0 (6.4) 84.0 (8.6)

ethyl 7.8 (8.5) 90.3 (7.3) 82.8 (13.2)

ethyl 4.6 (15.7) 104.1 (8.3) 73.9 (15.7)

7.8 (11.0) 97.4 (9.0) 96.2 (14.8)8.7 (5.6) 89.4 (8.5) 98.6 (5.7)9.7 (6.2) 91.7 (6.5) 73.7 (10.3)

11.1 (14.9) 94.1 (15.1) 74.2 (13.9)3.0 (16.9) 80.0 (10.3) 76.7 (6.9)

13.1 (14.0) 86.9 (8.1) 77.7 (12.7)10.4 (11.1) 92.3 (6.1) 79.2 (12.4)8.2 (5.1) 105.8 (5.7) 93.7 (11.8)

17.9 (10.2) 100.0 (9.0) 96.9 (15.0)11.1 (6.4) 93.2 (4.8) 72.4 (15.6)6.2 (8.4) 95.8 (9.3) 96.1 (15.7)

12.3 (8.0) 104.9 (9.4) 87.5 (9.1)15.5 (10.0) 91.5 (8.5) 80.4 (9.3)7.7 (3.7) 96.9 (11.8) 93.1 (8.1)

13.1 (17.2) 91.7 (12.4) 90.3 (15.8)9.4 (13.9) 88.3 (7.0) 79.8 (4.7)

14.4 (12.2) 92.8 (6.5) 81.9 (4.5)15.0 (9.3) 103.6 (9.8) 98.5 (6.5)14.5 (4.9) 102.5 (7.5) 101.2 (7.7)20.8 (12.7) 85.2 (4.1) 91.5 (14.8)25.5 (11.7) 98.1 (6.5) 102.0 (6.0)28.0 (8.5) 106.7 (8.3) 94.6 (7.0)9.3 (12.3) 93.8 (6.9) 84.6 (11.1)

17.9 (9.3) 99.4 (5.4) 93.8 (6.4)17.5 (7.0) 106.3 (8.4) 97.7 (7.3)3.9 (7.4) 93.3 (9.4) 87.2 (4.7)

21.2 (11.5) 109.9 (8.9) 97.7 (11.9)21.9 (10.1) 100.0 (6.1) 98.5 (10.5)24.8 (10.0) 105.6 (9.7) 93.0 (13.8)20.2 (11.4) 82.7 (5.7) 76.4 (12.0)17.7 (16.7) 86.1 (9.1) 93.2 (6.2)21.6 (9.8) 83.5 (10.8) 78.2 (12.5)

yl 27.7 (4.3) 99.5 (7.9) 106.4 (5.1)18.0 (14.0) 89.7 (4.7) 76.6 (7.1)27.9 (9.9) 102.0 (5.4) 93.5 (8.8)27.2 (8.3 88.3 (5.9) 83.2 (8.4)

action efciency. Relative standard deviation is given in brackets (n=5).andard deviation (RSD) is given in brackets (n=3).

elative recoveries determined when a white wine wasresentative matrix. No signicant difference betweent matrices is observed, indicating that the selectednothavea signicant inuenceon theextractionproce-rmore, recovery ranged from 72.7 to 112.0%, indicatingty of the extraction procedure in the checked matrices.that the analytical procedure does not depend on thealuatedand theHF-LPMEdevicesprovides clearextractstrix effect during UHPLCMS/MS determination.

ion

mized HF-LPME method was validated in terms of lin-racy, precision, selectivity and lower detection limits.

Identicwindows (Rstandard dewine samplconsecutivetwo MS/MSSelectivity osamples. Thtarget pestimay give a

Linearitysampleswiprocedure wgood linearSparkle wine Non-alcoholic beer Beer

109.4 (6.1) 102.2 (4.0) 93.9 (6.5)112.0 (7.7) 104.0 (5.6) 111.6 (12.7)80.4 (11.5) 71.0 (8.4) 75.9 (4.6)85.6 (9.6) 71.2 (13.9) 73.0 (11.5)

108.3 (5.6) 90.0 (8.9) 91.7 (13.4)90.2 (6.4) 106.1 (13.5) 109.0 (8.3)

109.7 (4.8) 108.6 (15.9) 95.1 (8.8)79.1 (10.9) 90.0 (10.9) 80.9 (9.1)82.9 (7.9) 82.4 (14.7) 75.3 (11.5)73.0 (8.1) 75.1 (12.0) 76.2 (10.2)

103.8 (12.2) 98.8 (5.5) 103.8 (8.6)108.8 (11.1) 111.2 (11.6) 98.0 (15.2)108.0 (10.8) 104.0 (4.0) 90.0 (11.0)93.1 (12.6) 80.0 (11.4) 88.2 (12.3)

86.5 (12.6) 103.5 (8.3) 76.1 (7.6)

104.4 (8.9) 111.5 (5.7) 103.8 (11.7)109.2 (13.7) 90.3 (7.1) 98.2 (4.9)102.3 (5.0) 92.9 (4.4) 85.7 (12.3)87.4 (12.3) 78.6 (12.9) 74.4 (11.9)92.7 (6.8) 101.3 (12.8) 106.7 (11.8)87.9 (15.9) 85.6 (12.4) 83.2 (12.6)

109.0 (8.2) 100.6 (9.4) 109.4 (6.8)88.2 (9.9) 103.7 (4.4) 108.5 (6.1)89.7 (11.4) 101.6 (16.0) 96.9 (8.9)72.2 (7.5) 80.7 (11.8) 79.0 (6.9)

103.9 (10.1) 97.1 (6.6) 104.2 (13.7)75.4 (6.0) 87.2 (6.9) 97.6 (11.2)81.7 (12.5) 95.6 (9.0) 82.6 (4.7)

104.9 (15.4) 94.3 (5.8) 109.7 (5.9)110.0 (10.5) 104.3 (8.8) 106.3 (8.5)109.7 (5.1) 104.7 (6.0) 105.5 (6.0)91.4 (8.3) 87.2 (6.9) 87.9 (5.1)

109.5 (6.9) 110.1 (8.5) 102.4 (6.9)105.0 (6.1) 105.4 (4.0) 107.7 (8.0)87.1 (8.6) 93.3 (15.1) 82.9 (12.5)97.0 (7.4) 109.3 (15.0) 108.2 (12.9)98.5 (15.4) 93.8 (14.9) 101.2 (7.3)

103.6 (14.5) 101.6 (11.8) 109.5 (9.9)93.7 (8.0) 97.6 (15.9) 102.3 (13.6)99.5 (7.2) 108.0 (10.6) 106.2 (10.0)83.1 (13.0) 109.7 (7.8) 95.9 (7.2)91.0 (11.6) 87.6 (8.3) 102.7 (6.2)

107.6 (13.7) 106.7 (5.5) 103.5 (12.7)99.3 (10.2) 98.9 (12.9) 110.6 (10.8)71.2 (4.8) 70.3 (4.1) 71.4 (10.3)96.2 (7.1) 92.2 (13.7) 92.3 (4.0)87.9 (6.0) 80.6 (7.3) 80.3 (4.5)99.8 (15.3) 102.8 (10.9) 85.6 (11.5)80.2 (13.8) 72.7 (7.7) 84.4 (5.5)84.6 (6.0) 98.6 (10.9) 103.8 (10.2)81.7 (10.8) 78.2 (9.0) 82.0 (12.6)

ation of the pesticides was based on the retention timeTWs), dened as the retention time averages threeviations of the retention time when ten blank whitees spiked at 25g L1 were analyzed (Table 1) in vedays. The identitywas then conrmedby acquisition oftransitions, bearing in mind European guidelines [30].f the method was evaluated by running control blanke absence of any signal at the same retention time ascides indicated there were no matrix interferences thatfalse positive signal.of the method was tested by spiking blank white wine

thin the range from5 to 100g L1, before theHF-LPMEas applied. Peak area was selected as response and

ity was found for all pesticides at concentrations within

22 P.P. Bolanos et al. / J. Chromatogr. A 1208 (2008) 1624

Fig. 6. Compawine extracted

the tested ithan 0.95 (T

Precisionrepeatabilitspiking blaand 25g Linterdayprelyzed dailyrepeatabilitthan 20%, ecarb and trilow concenprocedure.

Limits ocalculatedconcentraticentrationa signal toeral, low LOfrom 0.03 (rison of a: (a) UHPLCMS/MS chromatogram of a spiked white wine (25g L1) injectedby the optimized HF-LPME procedure prior chromatographic separation.

nterval, with linear determination coefcients higherable 3).of the overall method was studied by performing

y and interday precision. Repeatability was evaluatednk white wine samples at two concentration levels (51), performing 5 replicates for each level (Table 3). Forcision, two spiked samples at 5 and25g L1 were ana-for a period of 5 days (Table 3). It can be observed thaty and interday precision, expressed as RSD, were lowerxcept for some pesticides such as simazine, iprovali-adimenol,which valueswere slightly higher than 20% attrations, indicating the good precision of the extraction

f detection (LODs) and quantication (LOQs) wereanalyzing spiked blank white wine samples at lowons. LODs and LOQsweredetermined as the lowest con-of the analytes that produce chromatographic peak atnoise ratio of 3 and 10, respectively (Table 3). In gen-Ds and LOQs were obtained. For instance, LOQ rangedfenazaquin and fenbuconazol) to 5.62g L1 (isopro-

turon), whi[9,12]. It shlegislationproducts suLOQ levelscommoditie

3.4. Analys

The appof 5 sampleof wine, incand white)of the resulquality contity controlcheck the rcalibration

The obtawere detecdirectly into the chromatographic system, and (b) the same spiked

ch were similar or lower than other reported resultsould be indicated that the MRLs xed by the Europeanare referred to wine grapes or cereals, not to derivatech as wine or beer; however it is worth noting thatare below the values established MRLs for this type ofs.

is of real samples

licability of the method was evaluated by the analysiss of beer (alcoholic and non-alcoholic) and 15 samplesluding a sparkling wine and 2 home-made wine (redprepared by local farmers. In order to assure the qualityts when the proposed method was applied, an internalrolwas carried out in every batch of samples. This qual-implies a spiked blank sample at 25g L1 in order toeliability of the proposed method for each pesticide, acurve and a blank reagent.ined results are shown in Table 4 and no pesticides

ted in the analyzed beers. However, some pesticides,

P.P. Bolanos et al. / J. Chromatogr. A 1208 (2008) 1624 23

Table 3Validation parameters of the optimized HF-LPME procedure

Pesticide R2 LOD (g L1) LOQ (g L1) Repeatabilitya

Simazine 0.9520 1.50 5.00Desethyl terbuthylazine 0.9820 0.10 0.30Carbaryl 0.9886 1.20 3.80Monolinuron 0.9567 2.00 5.00Chlorotoluron 0.9729 1.00 3.40Metobromuron 0.9970 1.90 5.30Atrazine 0.9661 1.10 3.80Metazachlor 0.9734 1.78 4.28Lenacil 0.9883 1.60 5.10Fensulfothion 0.9806 1.40 4.90Isoproturon 0.9973 1.69 5.62Diuron 0.9718 1.57 5.22Azaconazole 0.9742 0.71 2.35Iodosulfuron mBensulfuron mDiethofencarbSebuthylazinePropazineLinuronSpiroxamineMethiocarbTerbuthylazinePaclobutrazolFlutalonilPromecarbPrometrynPropyzamideTriazophosTepraloxidimIprovalicarbTriadimenolFenhexamideEpoxiconazoleTebutamMetolachlorFenbuconazolDiubenzuronCyprodinilThiazopyrFurmecycloxSpinosadBitertanolPencycuronTrioxystrobinTriumizoleClethodimCycloxydimFluazifop buthSethoxydimHexythiazoxFenazaquin

a Expressedb Interday p

Table 4Concentration

Pesticide

CarbarylSpiroxamineTriadimenolEpoxiconazoleTriumizoleFenazaquinethyl 0.9923 0.70 2.33ethyl 0.9889 1.50 5.00

0.9624 0.50 1.660.9666 1.00 3.000.9842 0.71 2.370.9736 0.38 1.280.9932 1.10 3.670.9954 0.78 2.600.9535 0.51 1.700.9827 1.00 3.330.9952 1.42 4.260.9962 1.70 5.670.9650 1.60 5.330.9702 1.50 5.000.9588 0.41 1.370.9948 1.50 5.000.9967 1.54 5.130.9523 1.00 3.330.9503 0.09 0.300.9910 0.15 0.500.9544 0.20 0.670.9873 0.42 1.400.9891 0.01 0.030.9549 0.70 2.330.9634 0.09 0.300.9639 0.03 0.100.9702 0.07 0.230.9951 0.11 0.370.9918 0.01 0.040.9799 0.03 0.100.9762 0.50 1.670.9764 0.05 0.180.9978 1.00 3.330.9994 1.40 4.20

yl 0.9787 0.09 0.300.9839 1.00 3.330.9889 0.80 2.670.9840 0.01 0.03

as relative standard deviation (RSD) of 5 replicates.recision is given in brackets as RSD (n=5).

of pesticides detected in analyzed samples

Pesticides concentration (g L1)

S1 S9 S10 S11 S13 S17 S19

138.716.4 6.0

8.5 49.2 163.61.9

0.40.3

mainly inseand triadimone samplepresents hisuggestingWhereas trthe MRL eswas higherues indicatestablish drect use ofepoxiconazlevels.5g L1 25g L1

15.2 (21.2)b 3.9 (12.0)6.7 (8.9) 4.0 (6.6)

12.9 (19.1) 8.1 (11.3)11.5 (15.0) 8.1 (13.3)10.1 (14.4) 5.8 (13.2)14.2 (15.9) 10.3 (13.0)9.6 (12.6) 4.9 (9.3)8.7 (10.4) 1.8 (4.5)9.5 (14.2) 4.7 (9.9)6.3 (14.4) 3.7 (8.9)

11.4 (16.2) 7.2 (12.8)7.9 (10.8) 7.4 (8.6)5.8 (12.3) 3.3 (8.4)12.7 (14.3) 6.1 (8.5)14.2 (18.9) 9.8 (14.6)8.3 (13.5) 6.6 (11.4)7.5 (16.8) 5.9 (18.5)6.3 (13.4) 4.0 (10.2)4.0 (12.7) 3.6 (10.9)7.2 (13.4) 5.2 (10.9)8.4 (10.2) 5.9 (9.0)

10.7 (18.9) 3.7 (8.4)9.2 (16.2) 6.5 (12.8)9.6 (15.0) 6.7 (8.0)

12.3 (13.2) 6.4 (12.9)9.7 (11.3) 5.3 (10.1)

12.4 (17.4) 5.1 (12.9)12.3 (14.5) 7.4 (10.9)15.0 (18.4) 7.7 (18.2)12.3 (20.5) 11.6 (13.2)11.5 (20.7) 4.5 (9.9)6.1 (9.3) 3.0 (8.2)6.5 (12.3) 4.8 (6.1)6.2 (14.3) 4.4 (7.3)5.9 (10.4) 4.1 (5.9)5.4 (5.6) 4.0 (6.3)

11.7 (14.8) 10.7 (11.2)4.3 (9.1) 3.3 (5.2)5.6 (10.0) 4.1 (9.1)6.5 (8.7) 4.6 (7.9)9.2 (10.9) 5.6 (9.3)8.9 (12.5) 4.8 (8.3)8.6 (12.3) 4.5 (7.4)9.4 (18.1) 7.5 (14.1)8.7 (17.6) 4.2 (8.7)

14.1 (14.5) 7.2 (9.4)14.3 (18.7) 10.4 (10.6)9.3 (12.2) 7.1 (10.9)

14.8 (19.6) 7.3 (11.2)16.8 (18.5) 6.6 (14.2)4.6 (7.6) 3.8 (6.1)

cticides and fungicides, such as spiroxamine, carbarylenol, were detected in a few wine samples. Only(S13), which corresponds to a home-made wine,

gh amounts of pesticides (carbaryl and triadimenol),that grapes have been treated with these compounds.iadimenol concentration was ten times lower thantablished for wine grapes [31], carbaryl concentrationthan the MRL established for wine grapes. These val-e that more work should be carried out in order toenitive MRLs in wines in order to assure the cor-pesticides in this type of crops. On the other hand,

ole, triumizole and fenazaquin were detected at trace

24 P.P. Bolanos et al. / J. Chromatogr. A 1208 (2008) 1624

Fig. 7. UHPLCshowing quanwine containin0.4g L1 and

Finally,three posit138.7, 0.4observing tclean-up ef

4. Conclus

A new m51 pesticidmethod is vwhen extrabines the adpesticides, wsimplicity adisposableup proceduallows theholic beverprocedure i

operator skill should be high in order to get reproducible results.Despite of extraction time could be considered too long, a largenumber of samples can be extracted simultaneously, increasing thecapacity of the method. Thus, 12 samples can be analyzed in lessthan 3h, including sample preparation and determination, so themethod could be used in routine analysis.

Acknowledgements

authovat

06-1nishalsoProg

nces

. Poultam. 2asa-Cmissexes

ards mopeanabras,ytyl6 (200imne.oto, YazawaTheand InnAGL20theSpaRRG isCierva

Refere

[1] M.ECon

[2] H. B[3] Com

AnnregEur

[4] P. C[5] T. H

105[6] J.J. J

547[7] T. G

NakMS/MS extracted chromatograms of three positive real samplestication (upper) and conrmation (lower) transitions: (a) whiteg carbaryl at 138.7g L1, (b) white wine containing triumizole at(c) red wine containing fenazaquin (0.3g L1).

Fig. 7 shows the UHPLCMS/MS chromatograms forive results of carbaryl, triumizole and fenazaquin atand 0.3g L1, respectively in three wine samples,he high selectivity of the procedure due to the highciency of HF-LPME coupled with UHPLCMS/MS.

ion

ethod is proposed for the simultaneous extraction ofes in wine and beer using HF-LPME. Microextractionery selective and negligible matrix effects were foundcts were analyzed by UHPLCMS/MS. The method com-vantages of UPLCMS/MS, to separate and identify theith HF-LPME in terms of reduction of organic solvents,

nd elimination of carry over effect through the use ofmembranes. so HF-LPME can be used as an easy clean-re of complex matrices. The sensitivity of the methoddetermination of the pesticides at trace levels in alco-ages such as wine and beer. However, the extractions not automated and one of the main drawbacks is the

[8] J.W.B. BraPoppi, J. C

[9] C. MolinaDelgado,

[10] O. Nnez[11] A. Kruve,[12] J.W. Won

S.E. Ebele[13] S. Milln,

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A 942 (20[15] Y. Hayasa

Chem. 37[16] K.J. Chia,[17] M. Schell[18] D.A. Lamb[19] S. Pederse[20] K.E. Rasm[21] E. Psillaki[22] R. Romer

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Directiveinterpreta17, 2002,

[31] European90/642/Eat http:/2005.ors gratefully acknowledge Spanish Ministry of Scienceion (MICINN-FEDER) for nancial support (Project Ref.2127-C02-01). PPB acknowledges her grant (F.P.U.) fromMinistry of Science and Innovation (Ref. AP2005-3800).grateful for personal funding through the Juan de la

ram (Spanish Ministry of Science and Innovation-EFS).

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. Ito, H. Oka, I. Saito, H. Matsumoto, H. Sugiyama, C. Ohkubo, H., H. Nagase, Anal. Chim. Acta 531 (2005) 79.ga, C.B.G. Bottoli, I.C.S.F. Jardim, H.C. Goicoechea, A.C. Olivieri, R.J.hromatogr. A 1148 (2007) 200.-Mayo, J. Hernndez-Borges, T.M. Borges-Miquel, M.A. Rodrguez-J. Chromatogr. A 1150 (2007) 348., E. Moyano, M.T. Galcern, Trends Anal. Chem. 24 (2005) 683.A. Knnapas, K. Herodes, I. Leito, J. Chromatogr. A 1187 (2008) 58.g, M.G. Webster, D.Z. Bezabeh, M.J. Hengel, K.K. Ngim, A.J. Krynitsky,r, J. Agric. Food Chem. 52 (2004) 6361.M.C. Sampedro, N. Unceta, M.A. Goicolea, E. Rodrguez, R.J. Barrio, J.gr. A 995 (2003) 135.ro, C. Yage-Ruiz, B. Cancho-Grande, J. Simal-Gndara, J. Chromatogr.02) 41.ka, K.MacNamara, G.A. Baldock, R.L. Taylor, A.P. Pollnitz, Anal. Bioanal.5 (2003) 948.S.D. Huang, Rapid Commun. Mass Spectrom. 20 (2006) 118.in, B. Hauser, P. Popp, J. Chromatogr. A 1040 (2004) 251.ropoulou, T.A. Albanis, J. Biochem. Biophys. Methods 70 (2007)195.n-Bjergaard, K.E. Rasmussen, J. Chromatogr. A 1184 (2008) 132.ussen, S. Pedersen-Bjergaard, Trends Anal. Chem. 23 (2004) 1.s, N. Kalogerakis, Trends Anal. Chem. 22 (2003) 565.o-Gonzlez, E. Pastor-Montoro, J. Martnez-Vidal, A. Garrido-Frenich,

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Application of hollow fibre liquid phase microextraction for the multiresidue determination of pesticides in alcoholic beverages by ultra-high pressure liquid chromatography coupled to tandem mass spectrometryIntroductionExperimentalReagents and materialsInstruments and apparatusSample preparation by HF-LPMEUHPLC-MS/MS analysisSamples

Results and discussionsOptimization of the HF-LPME parametersStudy of matrix effect on the extraction processValidationAnalysis of real samples

ConclusionAcknowledgementsReferences