LY∕T 2904-2017 英文版 沉香

ICS79.080

B70

LY

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.Incaseofanydoubtaboutthecontents

ofEnglishtranslation,theChineseoriginalshallbeconsideredauthoritative.

ThisstandardisdraftedinaccordancewiththerulesgivenintheGB/T1.1—2009Directives

forstandardization—Part1:Structureanddraftingofstandards.

ThisstandardwasproposedbytheEndangeredSpeciesImportandExportManagementOfficeof

thePeople’sRepublicofChina.

ThisstandardwaspreparedbySAC/TC41(NationalStandardizationTechnicalCommittee41on

Timber,StandardizationAdministrationofChina).

I

LY/T2904—2017

Introduction

Aquilariaspp.isincludedinAppendixIIofConventiononInternationalTradeinEndangered

SpeciesofWildFaunaandFlora(CITES),andAquilariasinensisislistedunderprotectionlevel

ⅡintheCatalogueoftheNationalProtectedKeyWildPlants(theFirstBatch)(1999)inChina.

Thisstandardisdevelopedtoconserveandsustainablyuseagarwoodresources,andtopromote

thehealthydevelopmentoftheagarwoodindustryinChina.

II

LY/T2904—2017

Agarwood

1Scope

Thisstandardspecifiesthetermsanddefinitions,requirements,testmethodanddetermination

ofagarwood.

Thisstandardisapplicabletotheinspectionandidentificationofagarwoodrawmaterials

andagarwoodproducts.

2Normativereferences

Thefollowingreferenceddocumentsareindispensablefortheapplicationofthisdocument.

Fordatedreferences,onlytheeditioncitedapplies.Forundatedreferences,thelatest

editionofthereferenceddocument(includinganyamendments)applies.

GB/T29894—2013Generalmethodofwoodidentification

LY/T1788—2008Standardterminologyrelationgtowoodproperties

3Termsanddefinitions

Forthepurposesofthisdocument,thetermsanddefinitionsgiveninLY/T1788—2008andthe

followingapply.

3.1

Aquilariaspecies

plantspeciesbelongingtotheAquilariagenusofThymelaeceaefamilyasperplanttaxonomy

3.2

Aquilariawood

xylemtissueofAquilariaspecies

3.3

agarwood

naturalmixturecomposedofxylemtissueanditssecretions,formedduringthegrowthoftree

ofAquilariaspecies

3.4

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LY/T2904—2017

ethanolextractivesofagarwood

substancesextractedfromagarwoodwith95%ethanol,mainlyincluding2-(2-phenethyl)

chromones,sesquiterpenes,aromaticcompoundsandfattyacids

3.5

referencesubstanceofagarwood

standardmaterialusedforidentification,test,andcomparisonbythin-layer

chromatography(TLC)andhighperformanceliquidchromatography(HPLC),preparedand

calibratedbyanationaldesignatedmetrologyorinspectionagency

4Requirements

Thexylemstructureandsecretionscharacteristicsofagarwoodshallconformtothe

requirementsgiveninTable1.

Table1Requirementsforxylemstructureandsecretionscharacteristicsofagarwood

Testitems

Requirements

Diffuseporous;growthringsareindistinct;the

axialparenchymaisusuallyabsentwithahandlens;

thenumberofraysaremedium,veryfinetoslightly

fine;thenumberoftheincludedphloemislarge,

visibletothenakedeye,distinctwithahandlens

Macrostructure

Mainlyradialmultiplevessels,simpleperforation

plates,alternateintervesselpitting;vessel-ray

pittingissimilartointervesselpitting;axial

parenchymaisextremelyrare,vasicentric;fibers

arethin-walled;raysaremostlyuniseriate,

occasionallybiseriate;raysaremostlyone)rowsof

squareoruprightmarginalcellshomogeneousand

uniseriate,withasmallnumberofheterogeneousIII

orIItype;includedphloemispresentinlarge

amounts,foraminateorislandtype

Xylemstructure

Microstructure

Ethanol

Secretions

≥10.0%

extractives

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LY/T2904—2017

Cherry-red,purpleblue,lightred,orlightpurple

Chromogenic

reaction

ispermitted;colorlessorlightyellowisnot

permitted

Fluorescentspotsshallappear,correspondingin

positionandcolortothoseinchromatogramof

agarwoodreferencesubstance

TLC

6characteristicpeaksshowninFigure1shallappear,

correspondingtothoseinchromatogramofagarwood

referencesubstance;retentiontimeofpeak1shall

beconsistentwiththatofagarwoodreference

substance

HPLC

characteristic

chromatogram

1(Agarotetrol)

2

345

6

0

10

20

30

40

50

60

Retentiontime/min

Figure1HPLCcharacteristicchromatogramofagarwood

5Testmethod

5.1Agarwoodxylemstructure

5.1.1Sampling

Samplingiscarriedoutfromperpendiculardirectiontothecross,radialandtangential

section.Generally,thesizeshallbenolessthan10mm×10mm×10mm.Whentherequirements

arenotmet,thesamplingsizeshouldnotbelessthan5mm×5mm×5mm.

5.1.2Macrostructurecharacteristics

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LY/T2904—2017

Observeandrecordthecolor,odor,texture,andstructurecharacteristicsofthewoodsample;

observethewoodsamplewiththenakedeyeorhandlensof10×magnification,andrecordthe

characteristicsofheartwood,sapwood,growthrings,pores,axialparenchyma,rays,and

includedphloemfromthecrosssectionofthesample.RefertoAnnexAforthemacrostructure

characteristicsofxylemofAquilariaspecies.

5.1.3Microstructurecharacteristics

5.1.3.1Softening

Accordingtotheprovisionsof5.2.1and5.2.2inGB/T29894-2013,thesamplesshallbesoftened

bytheboilingmethodortheglycerol-ethanolmethod.

5.1.3.2Preparingsections

Placethesoftenedsampleonamicrotomeandcuttransverse,radial,andtangentialsections

withathicknessof15~20µmrespectively;oruseasuitableknifetopreparesectionsby

hand.Themicroscopicsectionsshallbepreparedfollowingthestepsofstaining,dehydration,

clearingandmounting.

5.1.3.3Recordingmicroscopiccharacteristics

Themicroscopicsectionshallbeplacedunderalightmicroscope,andthemicroscopicfeatures

includingvessels,axialparenchyma,woodfibers,rays,andincludedphloemsshallbeobserved

andrecorded.RefertoAppendixAformicrostructurecharacteristicsofxylemofAquilaria

species.

5.2CharacteristicsofAgarwoodsecretions

5.2.1Sampling

5.2.1.1Apparatus

5.2.1.1.1Mill.

5.2.1.1.2Screen,24mesh(850±29μm).

5.2.1.1.3Balance,accurateto0.001g.

5.2.1.2Testingprocedure

Takeabout10gofrepresentativesample,grinduntiltheentireportionspassa24-mesh

screen,andmixthoroughly.Halfofthesampleisusedforanalysis,andtheotherhalfis

reserved.

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5.2.2Determinationofmoisture

5.2.2.1Reagentsandapparatus

5.2.2.1.1Phosphoruspentoxide,analyticalreagent.

5.2.2.1.2Petridish,12cmindiameter.

5.2.2.1.3Weighingbottle,5cmindiameter.

5.2.2.1.4Vacuumdesiccator,30cmindiameter.

5.2.2.1.5Dryingtubewithanhydrouscalciumchloride.

5.2.2.1.6Balance,accurateto0.0001g.

5.2.2.2Testingprocedure

Distribute0.5~1.0cmdepthofphosphoruspentoxideinapetridish,andputthedishinthe

vacuumdesiccator.

Placeacleanweighingbottleinthevacuumdesiccator,removethestopperofthebottle.Reduce

thepressureofthedesiccatorbysuctiontolessthan2.67kPaandkeepfor30min,Maintain

thevacuumatroomtemperaturefor24hours.Connectthedryingtubewithanhydrouscalcium

chloridetotheairoutlet,loosenthedesiccatorplungertoequalizetheairpressure.Wait

untiltheinsideairpressureandoutsidepressureisconsistent,switchofftheplunger,open

thedesiccator,fitthestopper,takeouttheweighingbottleandweighpromptly.

Weighduplicatesamplesof0.5~1.0gtoanaccuracyof0.0001g,putinthedriedweighing

bottle,dryandweighinthesamewayasabove.Caculatethemoisturecontentinsampleaccording

toformula(1).Carryouttwosimultaneousmeasurements.Theabsoluteerrorbetweenthetwo

measuredvaluesshouldnotexceed0.3%,andtheresultshallbeexpressedastheaverage

moistureofduplicatesamplestothenearest0.01%.

W（%）m1m2

100

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

ms

where

m1——originalsamplemassplusmassofweighingbottle,unitingrams(g);

m2——driedsamplemassplusmassofweighingbottle,unitingrams(g);

ms——originalsamplemass,unitingrams(g).

5.2.3Determinationofethanolextractivescontent

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LY/T2904—2017

5.2.3.1Reagentsandapparatus

5.2.3.1.195%ethanol,analyticalreagent.

5.2.3.1.2Conicalflask,250mL.

5.2.3.1.3Condenser.

5.2.3.1.4Pipette,25mLand100mL.

5.2.3.1.5Evaporatingdish,9cmindiameter.

5.2.3.1.6Desiccator,30cmindiameter.

5.2.3.1.7Balance,accurateto0.0001g.

5.2.5.1.4Temperature-controlleddryingoven,roomtemperature~200℃,accurateto0.1℃.

5.2.3.2Testingprocedure

Weighduplicatesamplesof2gtoanaccuracyof0.0001g,andputina250-mLconicalflask.

Whentherequirementisnotmet,thesamplesizeshouldbenolessthan0.5g.Add100mLof

95%ethanolwithpipette,fitthestopper,weighandallowtostandfor1h.Connectwithreflux

condenser,heattoboilingandsimmerfor1h.

Cool,removetheflask,stopperit,andweighagain.Add95%ethanoltorestoreitsoriginal

weight,shakethoroughlyandfilterwithfilterpaper.Measure25mLofthefiltratewitha

pipetteandtransferintoanevaporatingdishwhichhasbeenpreviouslydriedtoconstantweight.

Evaporatetodrynessonwaterbath,andthendryinanovenfor3hat103±2℃.Coolina

desiccatorfor30minandweighpromptly.Caculatethecontentofethanolextractivesinsample

accordingtoformula(2).Carryouttwosimultaneousmeasurements.Theabsoluteerrorbetween

thetwomeasuredvaluesshouldnotexceed0.3%,andtheresultshallbeexpressedastheaverage

ethanolextractivesofduplicatesamplestothenearest0.01%.

1

mm2

X（%）ms（1W）400.........................(2)

where

m1——massofethanolextractivesplusmassofevaporatingdish,unitingrams(g);

m2——massofevaporatingdish,unitingrams(g);

ms——originalsamplemass,unitingrams(g);

W——moisturecontentofsample,%.

5.2.4Chromogenicreaction

5.2.4.1Reagentsandapparatus

5.2.4.1.195%ethanol,analyticalreagent.

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LY/T2904—2017

5.2.4.1.237%concentratedhydrochloricacid,analyticalreagent.

5.2.4.1.3Vanillin,analyticalreagent.

5.2.4.1.4Alcoholburner.

5.2.4.1.5Watchglass,5cmindiameter.

5.2.4.1.6Evaporatingdish,9cmindiameter.

5.2.4.1.7Graduatedpipette,5mL.

5.2.4.2Testingprocedure

Take2~3mLfiltrateofethanolextractivespreparedin5.2.3totheevaporatingdish,heat

thebottomofevaporatingdishwithalcoholburneruntilthefiltrateisevaporatedtodryness,

covertheevaporatingdishwithawatchglasspromptly,andcontinuetoheatuntiloily

substancesappearonthewatchglass.Removethewatchglass,add1dropofconcentrated

hydrochloricacid,about0.05gofvanillin,and1~2dropsof95%ethanoltotheoilysubstances,

stand,andobservethecolorchange.

5.2.5Thin-layerchromatography

5.2.5.1Reagentsandapparatus

5.2.5.1.1Diethylether,analyticalreagent.

5.2.5.1.2Trichloromethane,analyticalreagent.

5.2.5.1.3Graduatedscale,themeasuringrangeis0~20cm,andtheminimumscaleisnomore

than0.5mm.

5.2.5.1.4Temperature-controlleddryingoven,roomtemperature~200℃,accurateto0.1℃.

5.2.5.1.5Balance,accurateto0.001g.

5.2.5.1.6Thin-layerplate,SilicagelG,usuallyactivatedat110℃for0.5hbeforeuse.

5.2.5.1.7Sampleapplicator,quantitativecapillary,manual,semi-automatic,or

full-automatic.

5.2.5.1.8Chromatographicchamber,glasschamberwithaflatbottomortwintroughanda

tightlyfittedlid.

5.2.5.1.9Detectiondevice,acameraobscuraequippedwithultravioletlight(UV)of365

nmandcorrespondingfilter.Additionalcameraequipmentcouldbeusedtotakepicture.The

lightsourceshouldhaveenoughintensityofillumination.

5.2.5.2Testingprocedure

Weigh0.2gofgroundagarwoodreferencesubstancetoanaccuracyof0.0001g,add30mLof

diethylether,ultrasonicateinawaterbathfor60min,thenfilter.Evaporatethediethyl

ethertodryness,anddissolvetheresiduein1mLoftrichloromethaneasreferencesolution.

Preparesamplesolutioninthesamemannerasthereferencesolution.

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LY/T2904—2017

Applyseparatelytheabovetwosolutionstothesameplatewithsampleapplicator.Thedistance

betweensamplezoneandloweredgeofthin-layerplateis1.5~2.0cm.Theappliedvolumeof

solutionisusually4μL,adjustedaccordingtotheseparationresolution.

Addaproperamountoftrichloromethane-ether(10:1)mobilephasetothechromatographic

chamber,placetheplateloadedwithsampleintothechromatographicchamber,keepthesolvent

levelabout5mmbelowthesamplezone,andcoverthechambertightly.Whenthemobilephase

movesovertheprescribeddevelopmentdistance,removetheplatefromthechamber,andallow

theplatetodry.Generally,8~15cmshallbedevelopedfornormalthin-layerplate,and5~8

cmforhighperformancethin-layerplate.

Examineunderultravioletlightat365nm,andcomparethechromatogramsofsamplewith

referencesolution.RefertoAnnexBforrepresentativeTLCchromatogramofagarwood.

5.2.6HPLCcharacteristicchromatogram

5.2.6.1Reagentsandapparatus

5.2.6.1.195%ethanol,analyticalreagent.

5.2.6.1.2Acetonitrile,chromatographicallypure.

5.2.6.1.3Formicacid,guaranteedreagent.

5.2.6.1.4Water,Grade1.

5.2.6.1.5Centrifugetubewithstopper,30mL.

5.2.6.1.6Pipette,10mL.

5.2.6.1.70.1%solutionofformicacid,preparedbeforeuse.Measure1mLofformicacidwith

apipette,makeupto1000mLwithwater,andshakethoroughly.Thesolutionshallbepassed

throughamembranefilter(poresize0.45µm).

5.2.6.1.8Balance,accurateto0.001g.

5.2.6.1.9Ultrasoniccleaner,withapowerof250Wandafrequencyof40kHz.

5.2.6.1.10HPLC,equippedwithaUVspectrophotometricdetectorandagradientelution

device.

5.2.6.1.11Chromatographiccolumn,DiamonsilC18orPhenomenexlunaC18(particlesize5μm,

columnlength25cm,innerdiameter4.6mm).

5.2.6.2Testingprocedure

Weigh0.2gofgroundagarwoodreferencesubstancetoanaccuracyof0.001g,putinstoppered

centrifugetube,add10mLof95%ethanolwithapipette,weigh,ultrasonicateinawaterbath

for1h,cool,andweighagain.Replenishthelossofweightwith95%ethanol,mixwell,stand,

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filterthesupernatantthrougha0.45-μmmembranefilter,anduseasreferencesolution.

Preparesamplesolutioninthesamemannerasthereferencesolution,ortake2mLfiltrate

ofethanolextractivespreparedin5.2.3,passthrougha0.45-μmmembranefilter,anduse

assamplesolution.

Chromatographicconditionsandsystemsuitabilityshallbeperformed.Useacetonitrileas

mobilephaseA,0.1%solutionofformicacidasmobilephaseB,eluteingradientat0.7mL

perminuteasspecifiedinTable2withcolumntemperature31℃andspectrophotometerset

at252nm.Thenumberoftheoreticalplatesofcolumnisnolessthan6000,calculatedwith

referencetothepeakofagarotetrol.

Table2Gradientelutionconditions

Time（min）

0～10

MobilephaseA（%）

MobilephaseB（%）

15→20

20→23

23→33

33

85→80

80→77

77→67

67

10～19

19～28

28～40

40～41

33→35

35

67→65

65

41～50

50.1～60

95

5

Injectseparately10μLoftheabovetwosolutionsintoHPLC,andcomparethechromatograms

ofsamplewithreferencesolution.RefertoAnnexCforrepresentativeHPLCchromatogramof

agarwood.

6Determination

Ifalltheresultsofxylemstructureandsecretionscharacteristicsconformtorequirements

giveninTable1,thesampleshallbeconsideredasagarwood.Ifoneinspectionitemfails,

thesampleshallnotbeconsideredasagarwood.

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AnnexA

(informative)

MaincharacteristicsofXyleminAquilariaspecies

Aquilariagenus（Thymelaeaceaefamily）

Foreigntradenames:Agarwood,Eaglewood

Treesanddistribution:Evergreentrees.Approx.22species;distributedinIndonesia,

Malaysia,Vietnam,Cambodia,Laos,Thailand,Myanmar,India,thePhilippines,Singapore,New

Guinea,Brunei,BhutanandChina.InChina,therearetwonativeAquilariaspecies－Aquilaria

sinensisandAquilariayunnanensis,mainlydistributedinGuangdong,Hainan,Guangxi,Yunnan,

andFujianprovince.

Macro-structuralfeatures(takeAquilariasinensisasanexample):Diffuse-porouswood.The

woodcolorisyellowishwhite.Oncethewoodisexposedtotheairforalongterm,itssurface

willturndark.Theheartwoodcolorandsapwoodcolorareindistinguishable.Thewoodisglossy

andhasamildfragrantandsweetodor;thereisnospecialtaste.Growthringsareindistinct,

andthereexistdarklinesbetweentherings.Thenumberofvesselsisrare,slightlysmall

tomedium,visiblewithahandlens.Thesizeofvesselsisconsistentandevenlydistributed

inadispersivearrangement;tylosesareabsent.Theaxialparenchymaisusuallyabsent.The

numberofraysaremedium,veryfinetoslightlyfine,andvisiblewithahandlens;there

areraystripesontheradialsection.Ripplemarksandintercellularcanalareabsent.The

numberofincludedphloemislarge,visiblewiththenakedeye,foraminateorislandtype,

distributedevenlyinthesecondaryxylem(FigureA.1).Thecolorofwherearomaticresinis

producedturnsdarker,andisyellowishbrownordarkbrown,inblacklinesorplaques.

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LY/T2904—2017

a)Longitudinalsectionofsolidwood

b)Crosssectionofsolidwood（12X）

FigureA.1Xylemmacrostructurepicture

Microstructuralcharacteristics(takeAquilariasinensisasanexample):Vesselsarecircular

toovalinoutlineasviewedincrosssection,mostly4to6/mm;theyareinradialmultiples

2

mainlyof2to4andinclusters,occasionallysolitary;diffuse;thediametersofmostvessels

arefrom85to135μm;tylosesareabsent.Simpleperforationswithslightlyinclined

perforationplate.Theintervesselpitsarealternate,vesturedwithincludedlenticular

apertures.Thevessel-raypitsaresimilartointervesselpitsinsizeandshape.Theaxial

parenchymacellsarescarceandvasicentric.Theyhavenodularendwallsthataredistinct.

Gumsandcrystalsareabsent.Thin-walledfibershavesimplepitswithnarrowborder;part

ofthesimplepitsareslightlycircular,withslit-likeorX-shapedapertures.Raysare

nonstoried,5to10/mm,mostlyuniseriatewithoccasionalbiseriaterays.Rayshave7to20

cellsinheight;raytissuesare