LY∕T 2904-2017 英文版 沉香

?ICS 79.080

LY

B70

ForestryIndustryStandardofthePeople’sRepublicofChina

LY/T2904—2017

Agarwood沉香

（EnglishTranslation）

Issuedate:2017-10-27 Implementationdate:2018-01-01

IssuedbyStateForestryandGrasslandAdministrationofthePeople'sRepublicofChina

LY/T2904—2017

Foreword

SAC/TC41isinchargeofthisEnglishtranslation.IncaseofanydoubtaboutthecontentsofEnglishtranslation,theChineseoriginalshallbeconsideredauthoritative.

ThisstandardisdraftedinaccordancewiththerulesgivenintheGB/T1.1—2009Directivesforstandardization—Part1:Structureanddraftingofstandards.

ThisstandardwasproposedbytheEndangeredSpeciesImportandExportManagementOfficeofthePeople’sRepublicofChina.

ThisstandardwaspreparedbySAC/TC41(NationalStandardizationTechnicalCommittee41onTimber,StandardizationAdministrationofChina).

I

庫七七標準下載

Introduction

Aquilariaspp.isincludedinAppendixIIofConventiononInternationalTradeinEndangeredSpeciesofWildFaunaandFlora(CITES),andAquilariasinensisislistedunderprotectionlevelⅡintheCatalogueoftheNationalProtectedKeyWildPlants(theFirstBatch)(1999)inChina.

Thisstandardisdevelopedtoconserveandsustainablyuseagarwoodresources,andtopromotethehealthydevelopmentoftheagarwoodindustryinChina.

II

Agarwood

1Scope

Thisstandardspecifiesthetermsanddefinitions,requirements,testmethodanddeterminationofagarwood.

Thisstandardisapplicabletotheinspectionandidentificationofagarwoodrawmaterialsandagarwoodproducts.

2Normativereferences

Thefollowingreferenceddocumentsareindispensablefortheapplicationofthisdocument.Fordatedreferences,onlytheeditioncitedapplies.Forundatedreferences,thelatesteditionofthereferenceddocument(includinganyamendments)applies.

GB/T29894—2013Generalmethodofwoodidentification

LY/T1788—2008Standardterminologyrelationgtowoodproperties

3Termsanddefinitions

Forthepurposesofthisdocument,thetermsanddefinitionsgiveninLY/T1788—2008andthefollowingapply.

3.13.1

Aquilariaspecies

plantspeciesbelongingtotheAquilariagenusofThymelaeceaefamilyasperplanttaxonomy3.2

Aquilariawood

xylemtissueofAquilariaspecies3.3

agarwood

naturalmixturecomposedofxylemtissueanditssecretions,formedduringthegrowthoftree

ofAquilariaspecies3.4

1

ethanolextractivesofagarwood

substancesextractedfromagarwoodwith95ethanol,mainlyincluding2-(2-phenethyl)chromones,sesquiterpenes,aromaticcompoundsandfattyacids

3.5

referencesubstanceofagarwood

standardmaterialusedforidentification,test,andcomparisonbythin-layerchromatography(TLC)andhighperformanceliquidchromatography(HPLC),preparedandcalibratedbyanationaldesignatedmetrologyorinspectionagency

4Requirements

ThexylemstructureandsecretionscharacteristicsofagarwoodshallconformtotherequirementsgiveninTable1.

Table1 Requirementsforxylemstructureandsecretionscharacteristicsofagarwood

Testitems

Requirements

Xylemstructure

Macrostructure

Diffuseporous;growthringsareindistinct;theaxialparenchymaisusuallyabsentwithahandlens;thenumberofraysaremedium,veryfinetoslightlyfine;thenumberoftheincludedphloemislarge,visibletothenakedeye,distinctwithahandlens

Microstructure

Mainlyradialmultiplevessels,simpleperforationplates,alternateintervesselpitting;vessel-raypittingissimilartointervesselpitting;axialparenchymaisextremelyrare,vasicentric;fibersarethin-walled;raysaremostlyuniseriate,occasionallybiseriate;raysaremostlyone)rowsofsquareoruprightmarginalcellshomogeneousanduniseriate,withasmallnumberofheterogeneousIIIorIItype;includedphloemispresentinlargeamounts,foraminateorislandtype

Secretions

Ethanolextractives

≥10.0

2

Chromogenicreaction

Cherry-red,purpleblue,lightred,orlightpurpleispermitted;colorlessorlightyellowisnotpermitted

TLC

Fluorescentspotsshallappear,correspondinginpositionandcolortothoseinchromatogramofagarwoodreferencesubstance

HPLC

characteristicchromatogram

6characteristicpeaksshowninFigure1shallappear,correspondingtothoseinchromatogramofagarwoodreferencesubstance;retentiontimeofpeak1shallbeconsistentwiththatofagarwoodreference

substance

2

35

46

1(Agarotetrol)

0 10 20 30 40 50 60

Retentiontime/min

Figure1HPLCcharacteristicchromatogramofagarwood

5Testmethod

5.1Agarwoodxylemstructure

5.1.1Sampling

Samplingiscarriedoutfromperpendiculardirectiontothecross,radialandtangentialsection.Generally,thesizeshallbenolessthan10mm×10mm×10mm.Whentherequirementsarenotmet,thesamplingsizeshouldnotbelessthan5mm×5mm×5mm.

5.1.2Macrostructurecharacteristics

3

Observeandrecordthecolor,odor,texture,andstructurecharacteristicsofthewoodsample;observethewoodsamplewiththenakedeyeorhandlensof10×magnification,andrecordthecharacteristicsofheartwood,sapwood,growthrings,pores,axialparenchyma,rays,andincludedphloemfromthecrosssectionofthesample.RefertoAnnexAforthemacrostructurecharacteristicsofxylemofAquilariaspecies.

5.1.3Microstructurecharacteristics

Softening

Accordingtotheprovisionsof5.2.1and5.2.2inGB/T29894-2013,thesamplesshallbesoftenedbytheboilingmethodortheglycerol-ethanolmethod.

Preparingsections

Placethesoftenedsampleonamicrotomeandcuttransverse,radial,andtangentialsectionswithathicknessof15~20μmrespectively;oruseasuitableknifetopreparesectionsbyhand.Themicroscopicsectionsshallbepreparedfollowingthestepsofstaining,dehydration,clearingandmounting.

Recordingmicroscopiccharacteristics

Themicroscopicsectionshallbeplacedunderalightmicroscope,andthemicroscopicfeaturesincludingvessels,axialparenchyma,woodfibers,rays,andincludedphloemsshallbeobservedandrecorded.RefertoAppendixAformicrostructurecharacteristicsofxylemofAquilariaspecies.

5.2CharacteristicsofAgarwoodsecretions

5.2.1Sampling

Apparatus

.1Mill.

.2Screen,24mesh(850±29μm).

.3Balance,accurateto0.001g.

Testingprocedure

Takeabout10gofrepresentativesample,grinduntiltheentireportionspassa24-meshscreen,andmixthoroughly.Halfofthesampleisusedforanalysis,andtheotherhalfisreserved.

4

5.2.2Determinationofmoisture

Reagentsandapparatus

.1Phosphoruspentoxide,analyticalreagent.

.2Petridish,12cmindiameter.

.3Weighingbottle,5cmindiameter.

.4Vacuumdesiccator,30cmindiameter.

.5Dryingtubewithanhydrouscalciumchloride.

.6Balance,accurateto0.0001g.

Testingprocedure

Distribute0.5~1.0cmdepthofphosphoruspentoxideinapetridish,andputthedishinthevacuumdesiccator.

Placeacleanweighingbottleinthevacuumdesiccator,removethestopperofthebottle.Reducethepressureofthedesiccatorbysuctiontolessthan2.67kPaandkeepfor30min,Maintainthevacuumatroomtemperaturefor24hours.Connectthedryingtubewithanhydrouscalciumchloridetotheairoutlet,loosenthedesiccatorplungertoequalizetheairpressure.Waituntiltheinsideairpressureandoutsidepressureisconsistent,switchofftheplunger,openthedesiccator,fitthestopper,takeouttheweighingbottleandweighpromptly.

Weighduplicatesamplesof0.5~1.0gtoanaccuracyof0.0001g,putinthedriedweighingbottle,dryandweighinthesamewayasabove.Caculatethemoisturecontentinsampleaccordingtoformula(1).Carryouttwosimultaneousmeasurements.Theabsoluteerrorbetweenthetwomeasuredvaluesshouldnotexceed0.3,andtheresultshallbeexpressedastheaveragemoistureofduplicatesamplestothenearest0.01.

W（%）=

m1-m2

ms

′100

.........................(1)

where

m1——originalsamplemassplusmassofweighingbottle,unitingrams(g);

m2——driedsamplemassplusmassofweighingbottle,unitingrams(g);

ms——originalsamplemass,unitingrams(g).

5.2.3Determinationofethanolextractivescontent

5

Reagentsandapparatus

.195ethanol,analyticalreagent.

.2Conicalflask,250mL.

.3Condenser.

.4Pipette,25mLand100mL.

.5Evaporatingdish,9cmindiameter.

.6Desiccator,30cmindiameter.

.7Balance,accurateto0.0001g.

.4 Temperature-controlleddryingoven,roomtemperature~200℃,accurateto0.1℃.

Testingprocedure

Weighduplicatesamplesof2gtoanaccuracyof0.0001g,andputina250-mLconicalflask.Whentherequirementisnotmet,thesamplesizeshouldbenolessthan0.5g.Add100mLof

95ethanolwithpipette,fitthestopper,weighandallowtostandfor1h.Connectwithrefluxcondenser,heattoboilingandsimmerfor1h.

Cool,removetheflask,stopperit,andweighagain.Add95ethanoltorestoreitsoriginalweight,shakethoroughlyandfilterwithfilterpaper.Measure25mLofthefiltratewithapipetteandtransferintoanevaporatingdishwhichhasbeenpreviouslydriedtoconstantweight.Evaporatetodrynessonwaterbath,andthendryinanovenfor3hat103±2℃.Coolinadesiccatorfor30minandweighpromptly.Caculatethecontentofethanolextractivesinsampleaccordingtoformula(2).Carryouttwosimultaneousmeasurements.Theabsoluteerrorbetweenthetwomeasuredvaluesshouldnotexceed0.3,andtheresultshallbeexpressedastheaverageethanolextractivesofduplicatesamplestothenearest0.01.

X（%）=

m1-m2

′400.........................(2)

ms′（1-W）

where

m1——massofethanolextractivesplusmassofevaporatingdish,unitingrams(g);

m2——massofevaporatingdish,unitingrams(g);

ms——originalsamplemass,unitingrams(g);

W——moisturecontentofsample, .

5.2.4Chromogenicreaction

Reagentsandapparatus

.195ethanol,analyticalreagent.

6

.237concentratedhydrochloricacid,analyticalreagent.

.3Vanillin,analyticalreagent.

.4Alcoholburner.

.5Watchglass,5cmindiameter.

.6Evaporatingdish,9cmindiameter.

.7Graduatedpipette,5mL.

Testingprocedure

Take2~3mLfiltrateofethanolextractivespreparedin5.2.3totheevaporatingdish,heatthebottomofevaporatingdishwithalcoholburneruntilthefiltrateisevaporatedtodryness,covertheevaporatingdishwithawatchglasspromptly,andcontinuetoheatuntiloilysubstancesappearonthewatchglass.Removethewatchglass,add1dropofconcentratedhydrochloricacid,about0.05gofvanillin,and1~2dropsof95ethanoltotheoilysubstances,stand,andobservethecolorchange.

5.2.5Thin-layerchromatography

Reagentsandapparatus

.1Diethylether,analyticalreagent.

.2Trichloromethane,analyticalreagent.

.3Graduatedscale,themeasuringrangeis0~20cm,andtheminimumscaleisnomorethan0.5mm.

.4Temperature-controlleddryingoven,roomtemperature~200℃,accurateto0.1℃.

.5Balance,accurateto0.001g.

.6Thin-layerplate,SilicagelG,usuallyactivatedat110℃for0.5hbeforeuse.

.7Sampleapplicator,quantitativecapillary,manual,semi-automatic,orfull-automatic.

.8Chromatographicchamber,glasschamberwithaflatbottomortwintroughandatightlyfittedlid.

.9Detectiondevice,acameraobscuraequippedwithultravioletlight(UV)of365nmandcorrespondingfilter.Additionalcameraequipmentcouldbeusedtotakepicture.Thelightsourceshouldhaveenoughintensityofillumination.

Testingprocedure

Weigh0.2gofgroundagarwoodreferencesubstancetoanaccuracyof0.0001g,add30mLofdiethylether,ultrasonicateinawaterbathfor60min,thenfilter.Evaporatethediethylethertodryness,anddissolvetheresiduein1mLoftrichloromethaneasreferencesolution.Preparesamplesolutioninthesamemannerasthereferencesolution.

7

Applyseparatelytheabovetwosolutionstothesameplatewithsampleapplicator.Thedistancebetweensamplezoneandloweredgeofthin-layerplateis1.5~2.0cm.Theappliedvolumeofsolutionisusually4μL,adjustedaccordingtotheseparationresolution.

Addaproperamountoftrichloromethane-ether(10:1)mobilephasetothechromatographicchamber,placetheplateloadedwithsampleintothechromatographicchamber,keepthesolventlevelabout5mmbelowthesamplezone,andcoverthechambertightly.Whenthemobilephasemovesovertheprescribeddevelopmentdistance,removetheplatefromthechamber,andallowtheplatetodry. Generally,8~15cmshallbedevelopedfornormalthin-layerplate,and5~8cmforhighperformancethin-layerplate.

Examineunderultravioletlightat365nm,andcomparethechromatogramsofsamplewithreferencesolution.RefertoAnnexBforrepresentativeTLCchromatogramofagarwood.

5.2.6HPLCcharacteristicchromatogram

Reagentsandapparatus

.195ethanol,analyticalreagent.

.2Acetonitrile,chromatographicallypure.

.3Formicacid,guaranteedreagent.

.4Water,Grade1.

.5Centrifugetubewithstopper,30mL.

.6Pipette,10mL.

.70.1solutionofformicacid,preparedbeforeuse.Measure1mLofformicacidwithapipette,makeupto1000mLwithwater,andshakethoroughly.Thesolutionshallbepassedthroughamembranefilter(poresize0.45μm).

.8Balance,accurateto0.001g.

.9Ultrasoniccleaner,withapowerof250Wandafrequencyof40kHz.

.10HPLC,equippedwithaUVspectrophotometricdetectorandagradientelutiondevice.

.11Chromatographiccolumn,DiamonsilC18orPhenomenexlunaC18(particlesize5μm,columnlength25cm,innerdiameter4.6mm).

Testingprocedure

Weigh0.2gofgroundagarwoodreferencesubstancetoanaccuracyof0.001g,putinstopperedcentrifugetube,add10mLof95ethanolwithapipette,weigh,ultrasonicateinawaterbathfor1h,cool,andweighagain.Replenishthelossofweightwith95ethanol,mixwell,stand,

8

filterthesupernatantthrougha0.45-μmmembranefilter,anduseasreferencesolution.Preparesamplesolutioninthesamemannerasthereferencesolution,ortake2mLfiltrateofethanolextractivespreparedin5.2.3,passthrougha0.45-μmmembranefilter,anduseassamplesolution.

Chromatographicconditionsandsystemsuitabilityshallbeperformed.UseacetonitrileasmobilephaseA,0.1solutionofformicacidasmobilephaseB,eluteingradientat0.7mLperminuteasspecifiedinTable2withcolumntemperature31℃andspectrophotometersetat252nm.Thenumberoftheoreticalplatesofcolumnisnolessthan6000,calculatedwithreferencetothepeakofagarotetrol.

Table2Gradientelutionconditions

Time（min）

MobilephaseA（）

MobilephaseB（）

0～10

15→20

85→80

10～19

20→23

80→77

19～28

23→33

77→67

28～40

33

67

40～41

33→35

67→65

41～50

35

65

50.1～60

95

5

Injectseparately10μLoftheabovetwosolutionsintoHPLC,andcomparethechromatogramsofsamplewithreferencesolution.RefertoAnnexCforrepresentativeHPLCchromatogramofagarwood.

6Determination

IfalltheresultsofxylemstructureandsecretionscharacteristicsconformtorequirementsgiveninTable1,thesampleshallbeconsideredasagarwood.Ifoneinspectionitemfails,thesampleshallnotbeconsideredasagarwood.

9

AnnexA(informative)

MaincharacteristicsofXyleminAquilariaspecies

Aquilariagenus（Thymelaeaceaefamily）Foreigntradenames:Agarwood,Eaglewood

Treesanddistribution:Evergreentrees.Approx.22species;distributedinIndonesia,Malaysia,Vietnam,Cambodia,Laos,Thailand,Myanmar,India,thePhilippines,Singapore,NewGuinea,Brunei,BhutanandChina.InChina,therearetwonativeAquilariaspecies－AquilariasinensisandAquilariayunnanensis,mainlydistributedinGuangdong,Hainan,Guangxi,Yunnan,andFujianprovince.

Macro-structuralfeatures(takeAquilariasinensisasanexample):Diffuse-porouswood.Thewoodcolorisyellowishwhite.Oncethewoodisexposedtotheairforalongterm,itssurfacewillturndark.Theheartwoodcolorandsapwoodcolorareindistinguishable.Thewoodisglossyandhasamildfragrantandsweetodor;thereisnospecialtaste.Growthringsareindistinct,andthereexistdarklinesbetweentherings.Thenumberofvesselsisrare,slightlysmalltomedium,visiblewithahandlens.Thesizeofvesselsisconsistentandevenlydistributedinadispersivearrangement;tylosesareabsent.Theaxialparenchymaisusuallyabsent.Thenumberofraysaremedium,veryfinetoslightlyfine,andvisiblewithahandlens;thereareraystripesontheradialsection.Ripplemarksandintercellularcanalareabsent.Thenumberofincludedphloemislarge,visiblewiththenakedeye,foraminateorislandtype,distributedevenlyinthesecondaryxylem(FigureA.1).Thecolorofwherearomaticresinisproducedturnsdarker,andisyellowishbrownordarkbrown,inblacklinesorplaques.

10

a)Longitudinalsectionofsolidwood b)Crosssectionofsolidwood（12X）

FigureA.1Xylemmacrostructurepicture

Microstructuralcharacteristics(takeAquilariasinensisasanexample):Vesselsarecirculartoovalinoutlineasviewedincrosssection,mostly4to6/mm2;theyareinradialmultiplesmainlyof2to4andinclusters,occasionallysolitary;diffuse;thediametersofmostvesselsarefrom85to135μm;tylosesareabsent.Simpleperforationswithslightlyinclinedperforationplate.Theintervesselpitsarealternate,vesturedwithincludedlenticularapertures.Thevessel-raypitsaresimilartointervesselpitsinsizeandshape.Theaxialparenchymacellsarescarceandvasicentric.Theyhavenodularendwallsthataredistinct.Gumsandcrystalsareabsent.Thin-walledfibershavesimplepitswithnarrowborder;partofthesimplepitsareslightlycircular,withslit-likeorX-shapedapertures.Raysarenonstoried,5to10/mm,mostlyuniseriatewithoccasionalbiseriaterays.Rayshave7to20cellsinheight;raytissuesaremostlyheterogeneousandwithoccasionallyheterogeneousIIIorIItype.