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METHOD ABSTRACTS205 / AMINOGLYCOSIDE ANTIBIOTICS IN FEEDAPRAMYCIN, GENTAMICIN, NEOMYCINAminoglycoside antibiotics are among those commonly used in animal feeds to manage intestinal microorganisms. Thebenecial effects include improved growth and generally healthier animal populations. Use of antibiotics creates a demandfor analytical procedures to verify concentrations in pre-mixes and feeds and in some instances for residue analysis in animalproducts.This note describes a simple, robust analytical method for the family of Aminoglycoside antibiotics in feeds and animalproducts. The sample is homogenized with a generic extraction solution and the crude soluble portion is directly injectedinto an HPLC ion-exchange column. The column efuent is then mixed with an OPA/Thiouor reagent which formshighly uorescent derivatives with the primary amine moieties of the antibiotics.Call if you need to analyze other Aminoglycosides.Extraction Procedure:Take one part feed:10 parts Extraction solution (w/v)Catalog No. 1700-1118 and homogenize for ve minutes.Centrifuge for 10 minutes. Three layers will form: the pellet,a supernatant emulsion and a soft layer of oating fat. Carefullylift the oating fatty layer with a spatula and discard.Transfer the emulsion to a sealable vial. Coagulate theemulsion by placing the vial in a boiling water bath for 15minutes. Centrifuge for 10 minutes. The clear supernate isltered (0.45 um Nylon) and placed in an autosampler vial.METHODAnalytical ConditionsColumn: ALKION cation-exchange, K+ form, 4 x 150 mm,Catalog No. 9410917ALKION Guard column, 3 x 20 mm,Catalog No. 9493020ApramycinNeomycinGentamicinComplexTemperature: 40 °CFlow Rate: 0.8 mL/minMobile Phase:1700-1101, Potassium buffer, K011700-1102, Potassium titrant, K021700-1103, Potassium ionic strength adjuster, K03Post-column ConditionsPost-column System: Pinnacle PCXReactor Volume: 0.15 mLTemperature: 45 °CReagent: o-Phthalaldehyde/Thiouor + Brij 35®Flow Rate: 0.3 mL/minDetection: Fluorometer ex: 330 nm, em: 465 nm“ALKION” and Thiofl uor are trademarks of Pickering Laboratories, Inc.“Brij 35” is a registered trademark of ICI Americas, Inc.05 10 15 20 25 30minCONDITIONS FOR SEPARATION OF APRAMYCINGENTAMICIN AND NEOMYCINTIME (Min) 1700-1101 % 1700-1102 % 1700-1103 %0 60 40 015 13.2 8.8 7815.1 12 10 7830 12 10 7830.1 0 22 7831 0 22 7831.1 60 40 037 60 40 0FLOW RATE: 0.8 mL/min,COLUMN TEMP.: 40 ºC,INJECTION VOLUME: 10 LREFERENCES:Driver, J. L., Thex, N., Raynie, D., Ofi tserova, M., Pickering, M. Single-Laboratory Validation forthe Quantifi cation of Neomycin B and Neomycin C in Animal Feeds by Liquid ChromatographyFluorescence Detection with Post-column Derivatization.(2009) J.AOAC Int, 92, 34-41
METHOD ABSTRACTSStandard obtainedfrom SigmaApramycinAPRAMYCIN ANALYSISTIME (Min) 1700-1101 % 1700-1102 % 1700-1103 %0 67 33 05 67 33 015 14.7 7.3 7820 14.7 7.3 7820.1 0 22 7821 0 22 7821.1 67 33 0SAMPLE: 11.1 nmole Apramycin on column010 20 30minStandard obtainedfrom SigmaGentamicin Complex{GENTAMICIN ANALYSISTIME (Min) 1700-1101 % 1700-1102 % 1700-1103 %0 43 31 2620 9 13 7830 9 13 7830.1 0 22 7831 0 22 7831.1 43 31 26SAMPLE: 5 x 10-6 g Gentamicin complex on column107010 20 30minStandard obtained fromUS PharmacopeiaNeomycinNEOMYCIN ANALYSISTIME (Min) 1700-1101 % 1700-1102 % 1700-1103 %0 60 40 015 13.2 8.8 7825 11 11 7825.1 0 22 7826 0 22 7826.1 60 40 0SAMPLE: 2.75 nmole Neomycin010 20 30minPRODUCT CATALOGPICKERING LABORATORIES10/11
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TABLE OF CONTENTSINTRODUCTION3 Cust
Page 6 and 7:
INTRODUCTIONINSTALLATION & TRAINING
Page 8 and 9:
“A LOT HAS CHANGED SINCE PRODUCIN
Page 10 and 11:
INSTRUMENTS & HARDWAREFEATURE HIGHL
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INSTRUMENTS & HARDWARE• Indoor us
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INSTRUMENTS & HARDWARECRX400 POST-C
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CHROMATOGRAMS, COLUMNS & GUARDSPick
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CHROMATOGRAMS, COLUMNS & GUARDSREVE
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CHROMATOGRAMS, COLUMNS & GUARDS70-M
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CHROMATOGRAMS, COLUMNS & GUARDSHIGH
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CHROMATOGRAMS, COLUMNS & GUARDSSTAN
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CHROMATOGRAMS, COLUMNS & GUARDSSODI
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CHROMATOGRAMS, COLUMNS & GUARDSHIGH
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CHROMATOGRAMS, COLUMNS & GUARDSSTAN
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CHROMATOGRAMS, COLUMNS & GUARDSCARB
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CHROMATOGRAMS, COLUMNS & GUARDSSEPA
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CHROMATOGRAMS, COLUMNS & GUARDSALKI
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CHROMATOGRAMS, COLUMNS & GUARDSCOND
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CHROMATOGRAMS, COLUMNS & GUARDSAFLA
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CHROMATOGRAMS, COLUMNS & GUARDSVOGL
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CHROMATOGRAMS, COLUMNS & GUARDSANAL
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“PURE CHEMICALS DO WHAT THEY ARE
Page 54 and 55:
REAGENTS & CHEMICALSSERAPREP & URIP
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REAGENTS & CHEMICALSRESTORECHROMATO
Page 58 and 59: REAGENTS & CHEMICALSCALIBRATION STA
Page 60 and 61: ELUANTSCHROMATOGRAPHIC GRADEPickeri
Page 62 and 63: APPLICATION KITSAmino Acid Analysis
Page 64 and 65: BECKMAN 6300/7300 COLUMNS & CHEMICA
Page 66 and 67: REPLACEMENT COMPONENTS, FITTINGS &
Page 74 and 75: “THE LCTECH STORY”Pickering Lab
Page 76 and 77: LCTECH / GPC PRODUCT LINEGPC ULTRA
Page 78 and 79: LCTECH / GPC PRODUCT LINEUtilizing
Page 80 and 81: LCTECH / GPC PRODUCT LINE• Can be
Page 82 and 83: LCTECH / GPC PRODUCT LINEFEATURE HI
Page 84 and 85: LCTECH / GPC PRODUCT LINEFEATURE HI
Page 86 and 87: LCTECH / GPC PRODUCT LINE3Intensity
Page 88 and 89: LCTECH / GPC PRODUCT LINEEVA IIICON
Page 90 and 91: LCTECH / MYCOTOXIN PRODUCT LINEUV D
Page 92 and 93: LCTECH / MYCOTOXIN PRODUCT LINESAMP
Page 94 and 95: LCTECH / MYCOTOXIN PRODUCT LINEOTAR
Page 96 and 97: LCTECH / MYCOTOXIN PRODUCT LINEACCE
Page 98 and 99: “CUSTOMERS REALLY ONLY WANT THE C
Page 100 and 101: METHOD ABSTRACTS203.1 / MULTI-RESID
Page 102 and 103: METHOD ABSTRACTS208 / MULTI-RESIDUE
Page 104 and 105: METHOD ABSTRACTS328 / ANALYSIS OF A
Page 106 and 107: METHOD ABSTRACTS382 / 70-MIN AMINO
Page 110 and 111: METHOD ABSTRACTS102 / ANALYSIS OF B
Page 112 and 113: METHOD ABSTRACTS120 / BROMATE ANALY
Page 114 and 115: METHOD ABSTRACTS108 / POST-COLUMN L
Page 116 and 117: METHOD ABSTRACTS112 / EXPANDED HPLC
Page 118 and 119: METHOD ABSTRACTS325 / ANALYSIS OF F
Page 120 and 121: METHOD ABSTRACTS206 continued from
Page 124 and 125: METHOD ABSTRACTS105.2 / PARALYTIC S
Page 126 and 127: METHOD ABSTRACTS104 / POLYETHER ANT
Page 128 and 129: METHOD ABSTRACTS124 / SULFONAMIDE A
Page 132 and 133: GENERAL TERMS & CONDITIONS1 CONTROL
Page 134: DISTRIBUTORSContinued from previous
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