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1 Irnin Journl of Phrmceuticl Reserch (2018), 17 (4): Received: Mrch 2016 Accepted: Decemer 2016 Originl Article Determintion of Enrofloxcin nd Ciprofloxcin Residues in Five Different Kinds of Chicken Tissues y Dispersive Liquid Liquid Microextrction Coupled with HPLC Njmeh Rezee Moghdm,, Seyed Rfie Arefhosseini, Afshin Jvdi c,d, Frzneh Lotfipur e, Msood Ansrin, Elnz Tmizi c,e nd Mhoo Nemti c,e* Drug Applied Reserch Center, Triz University of Medicl Sciences, Triz, Irn. Deprtment of Biochemistry nd Diet therpy, Fculty of Nutrition nd Food Sciences, Triz University of Medicl Sciences, Triz, Irn. c Food nd Drug Sfety Reserch Center,Helth Mngement nd Sfety Promotion Reserch Institute, Triz University of Medicl Sciences,Triz, Irn. d Deprtment of Food Hygine, Fculty of Veterinry Medicine, Triz Azd Islmic University, Triz, Irn. e Deprtment. of Phrmceuticl nd Food Control, Fculty of Phrmcy, Triz University of Medicl Sciences, Triz, Irn. Astrct Contmintion of food producing nimls y veterinry drug residues, prticulrly quinolones, is n essentil issue in food sfety tht cuses incresing concern in consumers. The im of this study ws to investigte the occurrence of enrofloxcin nd its min metolite, ciprofloxcin, in chicken tissue smples slughtered in Triz, Irn. Totlly 250 smples including liver, muscle, gizzrd, hert, nd skin were studied. Dispersive liquid-liquid microextrction technique (DLLME) ws used s simple, high performnce, low-cost, nd fst smple pretretment method followed y high-performnce liquid chromtogrphy with UV detection for quntittive nlysis. The residues of enrofloxcin were detected nd quntified in 26 liver (52%) nd 10 skin (20%) smples nd ciprofloxcin residues were detected in 3 skin (6%) smples nd ccurtely determined in 15 liver (30%) smples; however they were not detected in gizzrd, hert, nd muscle smples. The results showed the ccumultion of enrofloxcin nd ciprofloxcin residues in chicken liver nd skin. Keywords: Enrofloxcin; Ciprofloxcin; Dispersive liquid liquid microextrction (DLLME); HPLC; Chicken. Introduction Enrofloxcin(1-cyclosporyl-6-fluoro-1, 4dihydro-4-oxo-7-[4ethyl-1-piperzinyl]- 3-quinoline croxylic cid) is synthetic fluoroquinolone ntimicroil gent which hs wide spectrum ctivity ginst Enteroctericee nd other Grm-negtive * Corresponding uthor: E-mil: nemtim@tzmed.c.ir cteri nd some ctivity ginst certin Grm positive cocci (1-3). Enrofloxcin is tken orlly in chicken, turkeys, pigs, nd cttle (with food, milk replcement nd/or in drinking wter), or dministered prenterlly through intrmusculr injection to pigs or sucutneous injection to cttle. Ciprofloxcin is the min metolite of enrofloxcin nd it ppers in vrious rtios in foodstuffs fter the dministrtion of enrofloxcin (4). As result of low ioniztion nd expressive
2 Enrofloxcin nd Ciprofloxcin Residues in chicken tissuse lipophilicity, like other fluoroquinolones, enrofloxcin is spred well nd fst through ll tissues of the orgnism (5, 6). Altough the ntimicroil mechnism of enrofloxcin is not compeletly understood, it is confirmed tht it is ctericidl gent tht inhiits the function of two enzymes including topoisomerse II nd topoisomerse IV. Topoisomerse II (DNAgyrse) is responsile for DNA repliction, which is essentil for mintining sphericl twist in DNA (6). Veterinry drugs re used on lrge scle s growth promoters or for the prevention nd therpy of infectious diseses in food producing nimls such s pigs, clves, poultry, nd fish ecuse of their good effectiveness (7-9). The ntimicroil properties of enrofloxcin show tht it hs dvntges for use in poultry. It is used for the tretment of common poultry infections such s mycoplsml infection, colicillosis, nd psteurellosis (10). Helth prolems might occur s result of excessive use of veterinry drugs in food producing nimls, ecuse most of these sustnces my hve some importnt toxic effects such s genotoxicity, crcinogenicity, immunotoxicity, or endocrine effects on consumers intking these sustnces (11). The presence of enrofloxcin resides in foodstuffs my cuse llergic rection in hypersensitive individuls nd could led to the incresed pthogen resistnce to clinicl drugs in humns; therefore, they my got importnt consequences for pulic helth (7). The enrofloxcin residue could enter the food supply nd chnge the ecology of the intestinl flor of consumers. It is lso prtilly metolized to its min metolite nd ciprofloxcin tht ffects the humn intestinl flor, s well (5). Moreover, fluoroquinolone ntimicroils cn cuse phototoxic skin rection in humns (12) nd chondrotoxic effects on young nimls (13) nd tendon rupture (14). Additionlly, presence of residul mounts of this ntiiotic could cuse serious difficulties for food processors in food fermenttion control (11). Therefore, to ensure humn food sfety, different countries nd regions set vrious mximum residue limits (MRLs) on fluoroquinolone residues nd some countries hve more rigid regultions thn others (7). Becuse of the complexity of iologicl solid mtrices, smple pretretment nd preconcentrtion re key points in the determintion of ntiiotics in different niml ody tissues (15). Nowdys, smple preprtion methods tht generte minimum toxic wste nd re more environmentlly friendly re introduced. These techniques re dispersive liquid liquid microextrction (DLLME) (16), supported liquid memrne (17), hollow fier supported liquid memrne (18), solid phse extrction (SPE) (7), pressurized liquid extrction (19) nd microwve-ssisted extrction (20). DLLME is reltively novel microextrction method which is introduced y Rezee et l. in 2006 s high-performnce nd powerful preconcentrtion technique. It is simple, quick nd in ccordnce with the green chemistry (20, 21). Different cteriologicl, chemicl, nd immuno-enzymticl methods hve een reported for the nlysis of fluoriquinolone residues in foodstuffs (4). Chemicl methods such s high performnce liquid chromtogrphy (HPLC) tht re used in severl lortories cn simultneously determine different quinolones, with lower limits of detection (LOD) (21). As recently ntiiotics nd veterinry drugs re widely used in food producing nimls tht cn cuse hrmful effects on humn helth, the min purpose of this study ws to determine the presence of enrofloxcin nd ciprofloxcin residues in chicken smples in Irn using powerful seprtion technique, such s HPLC, coupled with UV detector. In this study, DLLME methodology hs een proposed to increse smple preprtion throughput. Experimentl Regents Enrofloxcin nd ciprofoxcin were purchsed from Fluk Biochemic-Sigm Aldrich (Stein-heim, Germny). Chloroform, phosphoric cid nd sodium hydroxide were otined from Merck Co. (Drmstdt, Germny). Acetonitrile nd methnol were of HPLC grde nd purchsed from Duksn Pure Chemicls Co. (Gyeonggi-do, Kore). 1183
3 Rezee Moghdm N et l. / IJPR (2018), 17 (4): Chicken Smples Totlly 250 liver, muscle, gizzrd, hert, nd skin smples were collected from different ttoirs in Triz, Irn. Orgnic chickens were used s lnk smples nd these smples were nlyzed to ensure tht they re free of enrofloxcin nd ciprofloxcin. The chicken smples were preserved t -20 ºC for further nlysis. Enrofloxcin nd Ciprofoxcin Added Mterils Individul stock solutions of enrofloxcin nd ciprofloxcin were prepred t concentrtion of 100 µg/ml vi dissolving the ccurtely weighed mount of ech ntiiotic in cetonitrile nd working stndrd solutions with concentrtion of 10 µg/ml were prepred through diluting the pproprite mount of stock solution with distilled wter. Stndrd solutions were covered with luminum foil nd kept t 4 C. All spiked solutions were kept t room temperture for 1 h efore nlysis. Apprtus The nlyses were performed using KNAUER high performnce liquid chromtogrphic system consisting of degsser (Biotech model 2003,Onsl, Sweden), n isocrtic pump (K- 1000, Knuer, Berlin, Germny) nd n UV Vis detector (Knuer K-2005, Berlin, Germny). A perfectsil trget-c18 column (4.6 mm 250 mm, 5µm) ws used for seprtion. The column temperture ws set t 25 ºC nd the injection volume ws 20 µl; the moile phse mde up of cetonitrile nd phosphoric cid uffer (0.01 M, ph 3) (25:75% v/v) were utilized with flow rte of 1.0 ml/min nd detection ws crried out t 278 nm. A centrifuge (Pheonix, Germny), vortex mixer (Heidolph, UK), ph meter (Metrohm, Switzerlnd) nd n oil less piston vcuum pump (Kwke Airvc, Tiwn) were used s well. Smple preprtion Chicken liver, muscle, gizzrd, hert, nd skin smples were prepred using technique previously descried y Moem et l. (7), with minor modifictions. The smples were crushed using kitchen lender seprtely nd 5 g of ech homogenized smple ws weighted ccurtely nd trnseferd into 10 ml centrifuge tue. Then, 5 ml of 25 mm phosphoric cid:cetonitrile (30:70 v/v) solution ws dded to the smple nd shken for 30 s. The smples were centrifuged for 10 min t 4500 rpm t room temperture. The superntnt ws filtered through 0.45 µm memrne filter nd trnsferred into test tue. In skin smples the ccumulted ft content t the top of the test tue ws seprted first nd then the smple ws filtered. The ph vlue of cetonitrile extrct ws djusted to 7.0 using NOH 0.1 N, to otin the highest extrction efficiencies. 1 ml of the cetonitrile extrct ws used for DLLME procedure. DLLME procedure According to the previously reported work y Moem et l. (7), the DLLME-HPLC-UV method ws done s the following: 5 ml of doule distillied wter ws trnsferred into screw-cp glss test tue with conicl ottom. After tht, 1.0 ml of disperser solvent (the cetonitrile extrct ) ws dded nd 200 µl of extrcting solvent (chloroform) ws injected rpidly into the mixture. The ternry component solvent system ws mixed immeditely y vortex mixer for 30 s. Then, the resulted cloudy solution ws centrifuged t 4500 rpm for 5 min nd the sedimented phse, lden with enrofloxcin nd ciprofloxcin, ws trnsferred into the microtue nd dried t 25 C under gentle strem of nitrogen gs. Finlly, the residue ws redissolved in 100 µl moile phse nd injected into the HPLC system. Method vlidtion Since, the previously reported method (7) hs een utilized for the nlysis of smples, ccording to the FDA guidelines on the vlidtion of ionlyticl methods (22), the method ws prtilly vlidted in terms of linerity, ccurcy, repetility, limit of detection (LOD), nd limit of quntifiction (LOQ). In order to evlute linerity of the method for different smples, seven point mtrix mtched clirtion curves were otined through spiking different lnk smples with enrofloxcin nd ciprofloxcin in the concentrtion rnge of 5 to 500 µg/kg. To illustrte, spiked smple with 1184
4 Enrofloxcin nd Ciprofloxcin Residues in chicken tissuse Tle 1. Anlyticl performnce prmeters for determintion of enrofloxcin nd ciprofloxcin in chicken liver. LOQ (μg/kg) LOD (μg/kg) Correltion coefficient (r) Clirtion eqution Dt point Liner rnge (μg/kg) Smple y = x CIP - Liver y = x ENR - Liver y = x CIP - Skin y = x ENR - Skin CIP: ciprofloxcin ENR: enrofloxcin concentrtion of 30 µg/kg ws prepred y dding 15 µl of working stndrd solutions to 5 g of lnk tissue smples; other concentrtions were lso prepred ccording to this method. LOD nd LOQ of the method for ech smple were clculted using the elow equtions: LOD = 3.3 SD/s LOQ = 10 SD/s Where s nd SD were the slope nd stndrd devition of the y-itercept of three individul clirtion curves (23). Accurcy nd precision studies were crried out using spiked smples with concentrtions t the lower, middle, nd upper levels of the linerity rnge. Ech spiked smple ws nlyzed using the method in triplicte nd the experimentlly derived concentrtions were clculted using the otined pek res nd clirtion eqution. The ccurcy nd repetility of the method were expressed s the percentge of the experimentlly derived concentrtion to the nominl concentrtion nd reltive stndrd devition (RSD%) of the clculted concentrtions, respectively. Recovery clcultios were done using the spiked smples with the concentertions covering the liner rnge. The otined recoveries were reported s the percentge of the recovered concentrtion using the DLLME-HPLC method to the known concentrtion which ws dded to spike the lnk smples. And finlly, in order to evlute the suitility of the HPLC method for the sumultnous nlysis of enrofloxcine nd ciprofloxcine, system suitility prmeters including resolution etween the peks, cpcity fctor, tiling fctor, nd numer of theoreticl pltes were clculted using the stndrd smples. Result nd Discussion Method vlidtion The linerity prmeters of the proposed DLLME-HPLC method hve een reported in Tle 1. As indicted in this tle, the method could detect nd quntify the little mounts of enrofloxcin nd ciprofloxcin in chicken smples, especilly in liver nd skin. Since the chieved LOQs were lower thn MRL vlue estlished y Europen Union Commission Regultion No 37/2010 (EU 37/2010), the method could e utilized to inspect these ntiiotics mounts in rel smples. According to the reporetd results in Tle 2, the method ws ccurte nd precise enough for the quntifiction of enrofloxcin nd ciprofloxcin in liver nd skin smples covering wide concentrtion rnge round the cceptle MRL vlue. The otined recoveries for spiked liver smples with concentrtions in the liner rnge were in the rnge of 93.7 ± 1.05 to ± 0.4 % for enrofloxcin nd 98.3 ± 0.6 to ± 0.9 % for ciprofloxcin nd the recoveries of spiked skin smples were etween 80.5 ± 1.2 % nd ± 7.1 % for enrofloxcin nd etween 82.2 ± 9.9 % nd ± 4.9 % for ciprofloxcin. Besides, s it cn e seen in Tle 3, the pplied HPLC method ws suitle for the intended propose owing to the cpcity fctor of more thn 1, tiling fctor of less thn 2 nd reolution fctor of more thn 1.5. It shoud e mentioned tht ll the lnk 1185
5 Rezee Moghdm N et l. / IJPR (2018), 17 (4): Tle 2. The ccurcy nd precision of the DLLME-HPLC method for the nlysis of enrofloxcin nd ciprofloxcin in chicken liver nd skin smples (n = 3). Repetility (RSD%) Accurcy (%) Concentrtion (μg/kg) Smple ± ± CIP - Liver ± ± ± ENR - Liver ± ± ± CIP - Skin ± ± ± ENR - Skin ± CIP: ciprofloxcin ENR: enrofloxcin smples were previously nlyzed to ensure the sence of quinolone residues; lso it is worth mentioning tht since the method could not detect the nlytes in the rel gizzrd, hert nd muscle smples nd consequently were not pplied to quntify them in mentioned smples, the method vlidtion results in these mtrices were not completely reported; however, the reltive recoveries of enrofloxcin nd ciprofloxcin were 84.0 ± 7.1 %, 83.0 ± 6.2 % in gizzrd smples, 93.0 ± 6.1 %, 90.0 ± 9.9 % in hert smples nd 97.0 ± 5.6 %, 94.0 ± 5.5 % in muscle smples. Rel smple nlysis The vlidted method ws pplied to determine the enrofloxcin nd ciprofloxcin residues in 50 liver, 50 hert, 50 muscle, 50 gizzrd nd 50 skin smples. Figure 1 shows the chromtogrms of liver smples. It should e mentioned tht since there is not ny significnt differences mong the otined chroromtogrms from the nlysis of different smples, only chromtogrm of liver smples were rought. According to the reported results in Tle 4, enrofloxcin residues were detected nd quntified in 26 liver (52%) nd 10 skin (20%) smples nd ciprofloxcin residues were determined in 15 liver (30%) smples. However, they were not detected in gizzrd, muscle, nd hert smples. It is worth sying tht the method detected ciprofloxcin residues in 3 Tle 3. System suitility prmeters of the proposed method for the quntifiction of enrofloxcin nd ciprofloxcin. Anlyte Cpcity fctor Tiling fctor Resolution fctor Theoreticl pltes numer Enrofloxcin (Retention time = 14.3 ± 0.4) Ciprofloxcin (Retention time = 10.9 ± 0.2)
6 Enrofloxcin nd Ciprofloxcin Residues in chicken tissuse Figure 1. The chromtogrms otined using the proposed DLLME-HPLC method; () stndrd solution with concentrtion of 1 µg/ml, () spiked liver smple with enrofloxin nd ciprofloxcin t concentrtion of 0.5 µg/ml, (c) rel liver smple continig enrofloxin nd ciprofloxcin. Pek identifiction: (1) ciprofloxcin, (2) enrofloxcin. skin (6%) smples, ut since their mounts were less thn clculted LOQ vlues, their exct concentrtions could not e reported. The Irnin Ntionl Stndrds Orgniztion hs not fixed MRL vlue for enrofloxcin in food smples. According to the EU 37/2010 document, the MRL ws estlished 200 µg/ kg for liver, 100 µg/kg for muscle nd 100 µg/ kg for skin nd ft (sum of enrofloxcin nd ciprofloxcin) (24). The sum of enrofloxcin nd ciprofloxcin residues were ove thn the MRL estlished y Europen Union in 28% of liver smples; however, the drug residues were elow the MRL in skin smples. Therefore, the otined results ttrct some ttention to the fct tht it is necessry to give scientific informtion to poultry reeders out the withdrwel period, durtion from the time ntiiotic dministered until it is legl to slughter the niml. Also it seems tht orgniztions which re responsile for the food qulity control must do more serious supervisions nd inspections. There re some reported studies out determintion of enrofloxcin nd ciprofloxcin in niml tissue smples s shown in Tle 5. A closer look t the tle indictes tht the pplied DLLME-HPLC method represented lower LOD mounts in comprison to the some other studies 1187
7 Rezee Moghdm N et l. / IJPR (2018), 17 (4): Tle 4. The results for determintion of enrofloxcin nd ciprofloxcin in rel chicken tissue smples. Enrofloxcin Ciprofloxcin Tissue Men(µg /kg) Rnge Tissue Men(µg /kg) Rnge Liver ± Liver 24.8 ± Skin 21.7 ± Skin NQ Muscle ND muscle ND Hert ND Hert ND* gizzrd ND gizzrd ND* NQ: Detected ut not quntified (< LOQ) ND: Not detected Tle 5. Comprision mong the differnt nlyticl methods utilized to determine enrofloxcin nd ciprofloxcin in solid smples. Method Mtrix Anlyte LOD(µg/ kg) Regression coefficient Recovery (%) Reference SPE-HPLC-UV Chicken muscle Enrofloxcin Ciprofloxcin (27) SPE-CE-MS Chicken muscle Enrofloxcin Ciprofloxcin (28) SPE-LC-UV c Chicken tissue Enrofloxcin Ciprofloxcin (25) SPE-LC-MS Chicken tissue Enrofloxcin Ciprofloxcin (25) SPE-CE-DAD d Chicken Enrofloxcin Ciprofloxcin (29) DLLME-LC-DAD e Swine muscle Enrofloxcin Ciprofloxcin (20) DLLME-LC-DAD Chicken liver Enrofloxcin Ciprofloxcin (7) DLLME-HPLC-UV Chicken liver Enrofloxcin Ciprofloxcin This study DLLME-HPLC-UV Chicken skin Enrofloxcin Ciprofloxcin This study Solid-phse extrction- high-performnce liquid chromtogrphy- ultrviolet. Solid-phse extrction- cpillry electrophoresis- mss spectrometry. c Solid-phse extrction- liquid chromtogrphy- ultrviolet. d Solid-phse extrction- cpillry electrophoresis- diode rry detection. e Dispersive Liquid liquid microextrction- liquid chromtogrphy- diode rry de 1188
8 Enrofloxcin nd Ciprofloxcin Residues in chicken tissuse which used SPE s preconcentrtion technique (25, 26). Additionlly, the utilized method resulted in lower LOD vlues compred to the previous works pplied DLLME in swine muscle (LOD of 16.4 µg/kg for enrofloxcin) (20) nd chicken liver (LODs of 5 nd 16 µg/kg for enrofoxcin nd ciprofloxcin, respectively) (7). Therefore, it cn e sid tht the present method could e utilized to detect lower mounts of enrofoxcin nd ciprofloxcin in chicken tissues, especilly in liver nd skin. Conclusion In the present study, DLLME-HPLC method ws successfully pplied for the extrction nd quntifiction of enrofloxcin nd ciprofloxcin in different tissues of chicken intended for humn consumption. This ws simple nd sensitive method with n dequte linerity, ccurcy, precision, nd recovery tht led to the significnt reduction in orgnic solvent consumption. All these dvntges mke the proposed method s n pproprite technique pplicle in the wide rnge of routine nlyticl lortories. Using this method enrofloxcin nd ciprofloxcin were found in some chicken tissues which rises the wreness out the need to consider stricter legisltion on the use of veterinry drugs in poultry in Irn. Acknowledgement This is pper of dtse from the thesis entitled Determintion of Enrofloxcin residue in chicken liver, hert, skin nd gizzrd smples registered in the Drug Applied Reserch Center. We lso grtefully cknowledge their help nd finncil ssistnce s grnt (project No.93.72). (1) (2) (3) (4) References Mrtindle. The Complete Drug Reference. S. S (Ed), Phrmceuticl Press, London, (2005). Mrtinez M, McDermott P nd Wlker R. Phrmcology of the fluoroquinolones: perspective for the use in domestic nimls. Vet. J. (2006) 172: Čupić V, Dorić S, Trilović D nd Pejčić Z. Antimicroil drugs in veterinry medicine. Veterinrski glsnik (2004) 58: Kiriš A, Mrinšek J nd Flj VC. Introduction of the HPLC method for the determintion of quinolone residues in vrious muscle tissues. Biomed. chrom. (2005) 19: (5) Chen T, Yun J, Feng X, Wei H nd Hu W. Effects of enrofloxcin on the humn intestinl microiot invitro. Int. J. Antimicro. Agents (2011) 37: (6) Člnjk E, Smjlović M, Čklovic F, Algić D, Čklovic K nd Smjlović A. Detection of enrofloxcin residues in chicken met y microiologicl (growth inhiition test) nd ELISA method fter experimentl prophylctic nd therpeutic ppliction. MESO: prvi. hrvtski čsopis o mesu (2011) 13: (7) Moem D, Nindi M nd Due S. Development of dispersive liquid liquid microextrction method for the determintion of fluoroquinolones in chicken liver y high performnce liquid chromtogrphy. Anl. Chim. Act (2012) 730: (8) Mesgri A. M, Nemti M, Bei H, Ansrin M nd Nourddgr A. Solid Phse Extrction nd Simultneous Determintion of Tetrcyclines Residues in Cttle Edile Tissues Using n HPLC-FL Method Irn. J. Prm. Res. (2012) 2: (9) Asi M.M Bei H, Ansrin M, Nourddgr A nd Nemti M. simultneous determintion of Tetrcyclines residues in Bovine milk smples y Solid phse extrction nd HPLC-FL method Adv. Phrm. Bull. (2011) 1: (10) Dimitrov D, Lshev L, Ynev S nd Pndov B. Phrmcokinetics of enrofloxcin in turkeys. Res. Vet. Sci. (2007) 82: (11) Toldrá F nd Reig M. Methods for rpid detection of chemicl nd veterinry drug residues in niml foods. Trends Food Sci. Tech. (2006) 17: (12) Kleck G, Urch F nd Urwyler H. Fluoroquinolone nticterils enhnce UVA-induced skin tumors. J. Photochem. Photoiol. B: Biology (1997) 37: (13) Sthlmnn R, Kühner S, Shkiei M, Schwe R., Flores J, Evnder S. nd Vn Sickle D. Chondrotoxicity of ciprofloxcin in immture egle dogs: immunohistochemistry, electron microscopy nd drug plsm concentrtions. Arch. Toxicol. (2000) 73: (14) Petrović J, Bltić M, Čupić V, Stefnović S nd Stojnović D. Residues of enrofloxcin nd its min metolite ciprofloxcin in roiler chickens. Act Vet. (2006) 56: (15) Ro GS, Rmesh S, Ahmd AH Tripthi HC Shrm LD nd Mlik JK. Phrmcokinetics of enrofloxcin nd its metolite ciprofloxcin fter intrmusculr dministrtion of enrofloxcin in gots. Vet. Res. Com. (2001) 25: (16) Chen H, Chen H, Ying J, Hung J nd Lio L. Dispersive liquid liquid microextrction followed y high-performnce liquid chromtogrphy s n efficient nd sensitive technique for simultneous determintion of chlormphenicol nd thimphenicol in honey. Anl. Chim. Act (2009) 632: (17) Msgti TAM nd Nindi MM. Multiresidue determintion of sulfonmides in vriety of iologicl 1189
9 Rezee Moghdm N et l. / IJPR (2018), 17 (4): mtrices y supported liquid memrne with high pressure liquid chromtogrphy-electrospry mss spectrometry detection. Tlnt (2004) 64: (18) Romero-González R, Frenich AG, Vidl JLM nd Aguiler-Luiz MM. Determintion of ochrtoxin A nd T-2 toxin in lcoholic everges y hollow fier liquid phse microextrction nd ultr high-pressure liquid chromtogrphy coupled to tndem mss spectrometry. Tlnt (2010) 82: (19) Jiménez V, Compnyó R nd Guiters J. Vlidtion of method for the nlysis of nine quinolones in eggs y pressurized liquid extrction nd liquid chromtogrphy with fluorescence detection. Tlnt (2011) 85: (20) Tsi WH, Chung HY, Chen HH, Hung JJ, Chen H.C, Cheng SH nd Hung TC. Appliction of dispersive liquid liquid microextrction nd dispersive microsolid-phse extrction for the determintion of quinolones in swine muscle y high-performnce liquid chromtogrphy with diode-rry detection. Anl. Chim. Act (2009) 656: (21) Yn H nd Wng H. Recent development nd pplictions of dispersive liquid liquid microextrction. J. Chrom. A (2013) 1295: (22) FDA. US Deprtment of Helth nd Humn Services. Guidnce for Industry Bionlyticl Method Vlidtion. USFDA (2013) (23) Nemti M, Vlizdeh H, Ansrin M nd Ghderi F. Development of simple nd sensitive HPLC method for determintion of glucosmine in phrmceuticl formultions. J. AOAC Int. (2007) 90: (24) Commission E. phrmcologiclly ctive sustnces nd their clssifiction regrding mximum residue limits in foodstuffs of niml origin. Off. J. Eur. Union (2010) 5: L 15 (25) Bilc S, Brrón D nd Bros J. New extrction procedure to improve the determintion of quinolones in poultry muscle y liquid chromtogrphy with ultrviolet nd mss spectrometric detection. Anl. Chim. Act (2006) 580: (26) Boyce MC. Determintion of dditives nd orgnic contminnts in food y CE nd CEC. Electrophoresis (2007) 28: (27) Christodoulou EA, Smnidou VF nd Ppdoynnis IN. Vlidtion of n HPLC-UV method ccording to the Europen Union Decision 2002/657/EC for the simultneous determintion of 10 quinolones in chicken muscle nd egg yolk. J. Chrom. B (2007) 859: (28) Jun-Grcí A, Font G nd Picó Y. Determintion of quinolone residues in chicken nd fish y cpillry electrophoresis-mss spectrometry. Electrophoresis (2006) 27: (29) Brrón D, Jiménez-Lozno E, Cno J nd Bros J. Determintion of residues of enrofloxcin nd its metolite ciprofloxcin in iologicl mterils y cpillry electrophoresis. J. Chrom. B: Biomed. Sci. Appl. (2001) 759: This rticle is ville online t
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