ProvisionalTranslationfromtheJapaneseOriginal

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MinistryofHealth,LabourandWelfareNotificationNo.394

TheMinisterofHealth,LabourandWelfarehaspartiallyrevisedtheSpecificationsandStandardsforFood,FoodAdditives,Etc.(MinistryofHealthandWelfareNotificationNo.370,1959),asgivenbelow,basedontheprovisionofParagraph1,Article11oftheFoodSanitationLaw.

June23,2006

JiroKawasaki

MinisterofHealth,LabourandWelfare

Item5(9)inSectionAGeneralCompositionalStandardsforFood,PartI

Foodshallberevisedasfollows:

(9)ChloramphenicolAnalyticalMethod

Apparatus

Liquidchromatograph-massspectrometer(LC/MSorLC/MS/MS)

Reagentsandtestsolutions

Inadditiontothereagentsandtestsolutionslistedbelow,usethoselistedinSectionCReagents/TestSolutions,Etc.,PartIIFoodadditives.Acetonitrile:Acetonitrileproducedforliquidchromatography.

Divinylbenzene-N-vinylpyrrolidineCopolymerCartridgeColumn(60mg):Useapolyethylenecolumnof12–13mmininnerdiameterpackedwith60mgofdivinylbenzen-N-vinylpyrrolidonecopolymer,oracolumnequivalenttothespecifiedoneinseparationcapability.

Divinylbenzene-N-vinylpyrrolidineCopolymerCartridgeColumn(200mg):Useapolyethylenecolumnof12–13mmininnerdiameterpackedwith200mgofdivinylbenzen-N-vinylpyrrolidonecopolymer,oracolumnequivalenttothespecifiedoneinseparationcapability.

WaterWaterproducedforliquidchromatography.

Referencestandard

Chloramphenicol:Thisproductcontainsnotlessthan99%chloramphenicol.

Decompositionpoint:208°C.

Procedure

Extraction

Honey

Weigh5.00gofthetestsample,previouslyhomogenized,anddissolveitin20mlofwater.

Royaljelly

Weigh1.00gofthetestsample,previouslyhomogenized,add60mlofmethanol/1%metaphosphoricacid(3:2),andhomogenizethemixtureagain.Filterthehomogenizedsamplebysuctionintoarotaryvacuumevaporatorthroughafilterpapercoveredwitha2-mmthicklayerofdiatomaceousearth.Washtheresidueonthefilterpaperwith15mlofmethanol/1%metaphosphoricacid(3:2),andfilterthewashingsbysuction.Combinethefiltratesintotheevaporator,andevaporatethemixtureto2mlat45oCorlower.

Foodsotherthanthosegiveninsectionsiandii

Weigh5.00gofthetestsample,previouslyground.Formuscle,thefatlayershouldberemovedasmuchaspossiblebeforegrinding.Tothesamplemeasuredout,add100mlofmethanol/1%metaphosphoricacid(3:2),andhomogenizethemixtureagain.Filterthehomogenizedsamplebysuctionintoarotaryvacuumevaporatorthroughafilterpapercoveredwitha2-mmthicklayerofdiatomaceousearth.Washtheresidueonthefilterpaperwith10mlofmethanol/1%metaphosphoricacid(3:2),andfilterthewashingsbysuction.Combinethefiltratesintotheevaporator,andevaporatethemixtureto30mlat45oCorlower.

Clean-up

Honey

Pour5mlofmethanoland5mlofwaterintoadivinylbenzene-N-vinylpyrrolidinecopolymercartridgecolumn(60mg),anddiscardtheeffluent.Pourthesamplesolutionobtainedbytheextractionmethoddescribedin4-a.intothecolumn,thenpour5mlof20%(vol)methanol.Discardtheeffluent.Pour6mlof60%(vol)methanol,andcollecttheeluateintoarotaryvacuumevaporator.Removethemethanolandwaterat45°Corlower.Dissolvetheresiduein1.0mlofacetonitrile/water(3:7),andusetheobtainedsolutionasthetestsolution.

Royaljelly

Pour10mlofmethanoland10mlofwaterintoadivinylbenzene-N-vinylpyrrolidinecopolymercartridgecolumn(200mg),anddiscardtheeffluent.Pourthesamplesolutionobtainedbytheextractionmethoddescribedin4-a.intothecolumn,thenpour4mlofwaterand4mlof5%(vol)methanolintothecolumn.Discardtheeffluent.Pour10mlof60%(vol)methanol,andcollecttheeluateintoarotaryvacuum

evaporator.Removethemethanolandwaterat45°Corlower.Dissolvetheresiduein1.0mlofacetonitrile/water(3:7),andusetheobtainedsolutionasthetestsolution.

Foodsotherthanthosegiveninsectionsiandii

Pour5mlofmethanoland5mlofwaterintoadivinylbenzene-N-vinylpyrrolidinecopolymercartridgecolumn(60mg),anddiscardtheeffluent.Pourthesamplesolutionobtainedbytheextractionmethoddescribedin4-a.intothecolumn,thenpour10mlofwater.Discardtheeffluent.Pour10mlofmethanol,andcollecttheeluateintoarotaryvacuumevaporator.Removethemethanolat40°Corlower.Dissolvetheresiduein1.0mlofacetonitrile/water(3:7),andusetheobtainedsolutionasthetestsolution.

Determination

Qualitativetests

Performqualitativetestsunderthefollowingconditions.Testresultsobtainedmustbethesameasthoseobtainedforthereferencestandard.Testingconditions

Columnpacking:Octadecylsilanizedsilicagel(2–5μminparticlesize)

Column:Astainlesssteeltube(2.0–6.0mmininnerdiameter,100–250mminlength)

Columntemperature:40°C

Mobilephase:Acetonitrile/10mmol/lammoniumformate(3:7).Theflo