新西兰双氰胺检测方法.docx

新西兰双氰胺检测方法.docx

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新西兰双氰胺检测方法

DeterminationofDicyandiamide,CyanuricAcidandMelamineinDairyProductsbyHighPerformanceLiquidChromatographyTandemMassSpectrometry（LC-MS/MS）

1.ConfidentialityStatement

ThismethodisthepropertyofAsureQualityLtd.andcannotbecopied,usedordistributedwithoutthewrittenpermissionoftheWellingtonLaboratoryBusinessManager.

2.SafetyandHealth

MSDsheetsofallchemicals,solventsandstandardsmustbeunderstoodbytheanalystbeforecommencingthemethod.Avoidinhalingvaporsorchemicals.Usetheassignedprotectiveclothingandequipmentatalltimes.Ensurethatyouarefamiliarwiththelocationoffireextinguishers,firstaidkitandanysafetyequipmentthatmayberequired.

RefertotheLaboratory'sHazardRegisterforSignificantHazardsidentifiedandthecontrolmethodsspecifiedtominimisethesehazards.

2.1SpecificHazardsandControlMethods

NotApplicable.

3.definitions

3.1ABBREVIATIONS

DCD

Dicyandiamide

HPLC

HighPerformanceLiquidChromatography

IR

IonRatio

IS

InternalStandard

KTP

KeyTechnicalPersonnel

LC-MS/MS

LiquidChromatographyTandemMassSpectrometry

LIMS

LaboratoryInformationManagementSystem

LOD

LimitofDetection

LOQ

Limit（orLevel）ofQuantification

LOR

LimitofReporting

M

MolecularMass

MRM

MultipleReactionMonitoring

MSDsheets

MaterialSafetyDatasheets

PVDF

PolyvinylideneFluoride

Q1

Quantificationion

Q2

Secondaryquantification（check）ion

QC

QualityControl

RR

RelativeResponse

RS

RecoveryStandard

RT

RetentionTime

SOP

StandardOperatingProcedure

U

UncertaintyofMeasurementwith95%confidenceinterval

WS

WorkingStandard

3.2DEFINITIONOFTERMS

Analyte:

Thisisthecomponentofasamplethathastobedetected,identifiedandquantified.Theterm“analyte”includes,whereappropriate,derivativesformedfromtheanalyteduringanalysis.

Analyticalsamples:

Samplesrequiredtobeanalysedquantitativelyforthepresenceoftargetanalytes.

ExternalStandard:

AsolutionintheHPLCsolventwhichhasbeenfortifiedwithknownamountsofanalytesandinternalstandard.

InternalStandard（IS）:

Compoundaddedatthebeginningoftheanalysis,whichissimilartothetargetanalytesinchemicalcompositionandbehaviour.

LimitofDetection（LOD）:

Thatconcentrationofanalytewhichgivesaninstrumentsignalsignificantlydifferentfromtheblank.Commonlydefinedasthemeanblanklevelplus3SDofblankmeasurements.

Limit（orLevel）ofQuantification（LOQ）:

Regardedasthelowerlimitforprecisequantitativemeasurements（Commonlydefinedaseitherthemeanblanklevelplus10SDofblankmeasurementsor3LOD.Commonlyisrepresentedbythelowestconcentrationcalibrationstandard,sothatsamplescannotbequantifiedbelowthisstandard（bydefinition））.

LimitofReporting（LOR）:

ThepracticallimitofresiduequantificationatorabovetheLOQ.TheLORmustbenolowerthanthelowestcalibrationstandard;alimitspecifiedbyaregulatorybodyorclient.

MatrixBlank（RecoveryBlank）:

Ablankpreparedfromthesametypeasthematrixofinterest.

MatrixRecoverySample（RecoverySample）:

Asamplepreparedfromthematrixblanktowhichisaddedknownamountsofanalytesandinternalstandardinordertoestablishtheefficacyofthemethod.Inthismethodtheyarealsousedascalibrationsamplestoquantifyanalytesinanalyticalsamples.

Reagentblankdetermination:

Thisisthecompleteanalyticalprocedureappliedwithomissionofthetestportionorusinganequivalentamountofsuitablesolventinplaceofthetestportion.

RelativeResponse（RR）:

Ratiooftheresponseofananalytetoitsinternalstandard.

RecoveryStandard（RS）:

Compoundaddedattheendoftheanalysis.

4.IntroductionandOverview

4.1INTRODUCTION

Foodcontaminantanalysisinagriculturalproductsisofvitalimportanceforthesafetyofourfoodandtosafeguardourexportmarkets.

4.2SCOPE

Themethodhasbeenvalidatedinmilkandmilkpowders.Itcanbeusedforothersimilarmatricesprovidedthatitisdemonstratedthatthemethodperformancevaluesaremet.Additionalvalidationandrecordsthatmayberequiredwillbeaccordingtotherelevantvalidationprocedureofthelaboratory.

NOTE:

Refertotherelevantvalidationprocedureforvalidationactionsrequiredifadditionalmatrices/analytestothosealreadyvalidatedarerequiredtobeusedbythemethod.

4.3METHODSUMMARY

Samplesofmilkorreconstituteddairypowderaredilutedwithacetonitrile,sodiumchlorideisaddedandaftercentrifugation,filtrationandadditionofarecoverystandardtheresultingacetonitrilesolutionisanalysedbyLC-MS/MS.

4.4ANALYTES

AnalytesandInternalStandardsaresummarisedinTable1.

TABLE1:

TestArticles（AnalytesandInternalStandards）

TestArticles

CASNumber*

Analytes

Dicyandiamide（2-cyanoguanidine）

461-58-5,M=84

CyanuricAcid

108-80-5,M=129

Melamine

108-78-1,M=126

InternalStandards

[13C315N3]Melamine（RecoveryStandard）

M=132

[13C3]Melamine

1173022-88-2,M=129

[13C3]CyanuricAcid

201996-37-4,M=132

*M=molecularmass

Structures

NotApplicable.

5.ReferencedDocuments

AM-006:

AnalyticalMethods–UnidentifiedAnalyticalResponses–InvestigationandRecordKeeping

AM-013:

Chemicals,ReagentsandSolutions

AsureQualityLimitedValidationReportVR-290,DeterminationofCyanuricAcidandMelamineinMilkandPowderedMilkproductsbyHighPerformanceLiquidChromatographyTandemMassSpectrometry（LC-MS/MS）,October2008

AsureQualityLimitedValidationReportVR-441,DeterminationofDicyandiamide,CyanuricAcidandMelamineinMilkandPowderedMilkproductsbyHighPerformanceLiquidChromatographyTandemMassSpectrometry（LC-MS/MS）,May2010

AsureQualityLimitedValidationReportVR-499,DeterminationofCyanuricAcidandMelamineinMilkandPowderedMilkproductsbyHighPerformanceLiquidChromatographyTandemMassSpectrometry（LC-MS/MS）.SupplementaryvalidationtoVR-290,January2011

EQ-004:

WorkingStandards–Preparation,CheckingandRecords

EQ-043:

EquipmentProcedureforLC-MS/MS

EQ-058:

PrimaryStandards–Ordering,StorageandRecords

QC-025:

UncertaintyofMeasurementinAnalyticalResults

QC-026:

ControlChartsProcedure

QC-028:

QualityControl–BlindSpike/Intra-LaboratoryCheckSampleScheme

QC-029:

QualityControlProceduresforAnalyticalResults

SR-012:

SamplePreparation

6.Reagents

RefertoAM-013.

DONOTUSEEXPIREDCHEMICALS,REAGENTSORSOLUTIONS.

Allreagentsandchemicalsmustbeofsuchagradethattheydonotinterferewiththeanalyticalprocess.Usethestatedgradeorequivalent.

6.1CHEMICALS

∙Aceticacid（analyticalgrade）

∙Acetonitrile,（pesticidegrade）

∙Ammoniumacetate（analyticalgrade）

∙Deionisedwater

∙Dichloromethane（analyticalgrade）

∙Sodiumchloride（analyticalgrade）

6.2SOLUTIONS

∙HPLCSolventA

Intoa1000mLSchottbottleadd0.77gofammoniumacetateand1000mLofdeionisedwater.Stiruntilthesolidisdissolved.

Storeatroomtemperature.Stablefor2weeks.

∙HPLCSolventB

Intoa2000mLSchottbottleadd1.54gofammoniumacetateand60mLofdeionisedwater.Stiruntilthesolidisdissolved.Usinga2000mLmeasuringcylinder,add1940mLofacetonitrile.Warmthebottletoapproximately50oC（waterbath）toaidcompletedissolutionofammoniumacetate.

Storeatroomtemperature.Stablefor2weeks.

7.STANDARDS

RefertoSOPsEQ-004andEQ-058.

Uponreceipt,anorange“WaitingApproval”labelisplacedonthestandard.OncethestandardhasbeenfoundtobeacceptableasdeterminedinEQ-058,theorangelabelisoverlaidbyagreen“QCReleased”label,datedandinitialled.

NOTE:

Donotuseexpiredprimarystandardsorstandardsolutions.Donotusestandardswhichhavenotbeenreleased/approvedforusebyaSeniorScientist,Scientist（approvedbytheTechnicalManager）ortheTechnicalManager.

7.1PRIMARYSTANDARDS

∙MelamineisavailablefromAcrosOrganics,>99%.Cataloguenumber22048

∙CyanuricacidisavailablefromAldrich98%.Cataloguenumber185809-25G

∙DicyandiamideisavailablefromAldrich,99%.Cataloguenumber76609

∙[13C315N3]Melamine（RecoveryStandard）isavailablefromCambridgeIsotopeLaboratories,100µg/mLinwater.CataloguenumberCNLM-8150-1.2

∙[13C3]Melamine（InternalStandard）VETRANAL,10mgisavailablefromSigma-Aldrich.Cataloguenumber32666-10MG.

∙[13C3]CyanuricAcid（InternalStandard）:

VETRANAL,10mg,isavailablefromSigma-Aldrich.Cataloguenumber32679-10MG,

7.2SECONDARYSTANDARDS

7.2.1UnlabelledMelamineandCyanuricAcidStandards（100µg/mL）

Weighaccurately10.0mgofstandardintoa125mLpolypropylenebottleandadd

100gofdeionisedwaterbyweighttogivea100µg/mLsolution.Capandshakethoroughlytodissolvethecompound.

Storeatroomtemperature.Solutionsarestablefortwoyears.

7.2.2DicyandiamideStandard（1mg/mL）

Weighaccurately10.0mgofstandardintoa10mLcalibratedvolumetricflask.Adddeionisedwater,shaketodissolveandmakeuptovolume.Transfertoa20mLscrewcapvialandcaptightly.

Storeatroomtemperatureinthedark.Solutionisstableforsixmonths.

7.2.3RecoveryStandard（1µg/mL）

Remove1.2mLoftheprimarystandardsolutionof[13C315N3]melaminewithacalibratedHamiltonsyringeanddiluteto120mLina125mLpolypropylenepottlewithwatertogivea1µg/mLsolution.

Secondarystandardshaveanexpirydateoftwoyearsfromthedateofpreparationofsolutionortheexpirydateoftheprimarystandard,whicheverisearlier.Thesestandardsaretobestoredatroomtemperature.

7.2.4InternalStandards（100µg/mL）

Weighaccurately10.0mgofstandardintoa125mLpolypropylenebottleandadd

100gofdeionisedwaterbyweight