UGANDA STANDARD  
       US ISO 6865
First Edition
   2009 -09-04
 
Reference numbe r
  US ISO 6865:2000
 © UNBS 200 9 
  
Animal feeding stuffs — Determination of crude fibre content—
Method with intermediate filtration 
 US ISO 6865:2000  
 
© UNBS  2009 – All rights reserved Compliance with this standard does not, of itself confer immunity from legal obligations 
A Uganda Standard does not purport to include all necessary provisions of a contract. Users are 
responsible for its correct application  
© UNBS 2009  
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permission from UNBS. 
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Tel: 256 41 505 995 
Fax: 256 41 286 123 
E-mail: unbs@infocom.co.ug
   US ISO 6865:2000  
© UNBS  2009 - All rights reserved National foreword 
Uganda National Bureau of Standards (UNBS) is a parastatal under the Ministry  of Tourism, Trade and 
Industry established by the Act of Parliament of 198 3, of the Laws of Uganda.  UNBS is mandated to co-
ordinate the elaboration of standards and is  
(a) a member of International Organisation for Standardisation (ISO) and  
(b) a contact point for the WHO/FAO Codex A limentarius Commission on Food Standards, and  
(c) the National Enquiry Point on TBT/SPS Agr eements of the World Trade Organisation (WTO). 
The work of preparing Uganda Standards is carried out through Technical Committees. A Technical 
Committee is established to deliberate on standards in a gi ven field or area and consists of representatives of 
consumers, traders, academician s, manufacturers, governme nt and other stakeholders. 
Draft Uganda Standards adopted by the Technical Committ ee are widely circulated to stakeholders and the 
general public for comments. The committee reviews the comments before recommending the draft standards 
for approval and declaration as Uganda Standards by the National Standards Council. 
This Uganda Stand ard, US ISO 6865:2000, Animal feeding stuffs — Determination of crude fibre 
content — Method with intermediate filtration, is identical with and  has been reproduced from an International
Standard ISO 6865:2000,  Animal feeding stuffs — Determination of crude fibre content — Method with
intermediate filtration, and has been adopted as a Uganda Standard . 
Wherever the words, "International Standard" a ppear, they should be replaced by "Uganda Standard." Reference number
ISO6865:2000(E)
©ISO2000INTERNATIONAL
STANDARDISO
6865
Firstedition
2000-10-15
Animal feedingstuffs— Determinationof
crude fibre content— Methodwithintermediate filtration
Aliments des animaux — Déterminationde la teneur encellulose brute—
Méthode avec filtration intermédiaireISO 6865:2000(E)
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ii © ISO2000 –All rights reservedISO 6865:2000(E)
© ISO2000 –All rights reserved iiiForeword
ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies (ISO
member bodies). The work of preparing International Standards is normally carried out through ISO technical
committees. Each member body interested in a subject for which a technical committee has been established hasthe right to be represented on that committee. International organizations, governmental and non-governmental, inliaison with ISO, also take part in the work. ISO collaborates closely with the International ElectrotechnicalCommission(IEC) onall matters of electrotechnicalstandardization.
InternationalStandards are drafted in accordance with the rules givenintheISO/IEC Directives, Part3.
Draft International Standards adopted by the technical committees are circulated to the member bodies for voting.
Publicationas anInternationalStandardrequires approval byat least 75% of themember bodies casting a vote.
Attention is drawn to the possibility that some of the elements of this International Standard may be the subject of
patent rights. ISO shall not beheldresponsiblefor identifyinganyor allsuch patent rights.
International Standard ISO6865 was prepared by Technical Committee ISO/TC34, Food products , Subcommittee
SC 10,Animalfeedingstuffs .
Annex A of this International Standardis for informationonly.INTERNATIONAL STANDARD ISO 6865:2000(E)
© ISO2000 –All rights reserved 1Animalfeedingstuffs —Determinationofcrude fibre content —
Methodwithintermediate filtration
1 Scope
This International Standard specifies a method with intermediate filtration for the determination of the crude fibre
content. Amanualprocedure and asemi-automatic procedureare described.
Themethodis applicableto animalfeeding stuffs with acrudefibrecontent greater than10g/kg.
NOTE For animal feeding stuffs with a crude fibre content equal to or less than 10 g/kg, the method described in ISO6541
[7] maybe used.
This International Standard is also applicable to cereals and pulses.
2 Normative references
The following normative documents contain provisions which, through reference in this text, constitute provisions of
this International Standard. For dated references, subsequent amendments to, or revisions of, any of thesepublications do not apply. However, parties to agreements based on this International Standard are encouraged to
investigate the possibility of applying the most recent editions of the normative documents indicated below. For
undated references, the latest edition of the normative documents referred to applies. Members of ISO and IECmaintain registers of currentlyvalid InternationalStandards.
ISO3696:1987, Water for analytical laboratory use —Specification and test methods.
ISO6498:1998, Animalfeedingstuffs —Preparationof test samples.
3 Term and definition
For the purposes of this International Standard, thefollowing term anddefinition apply.
3.1
crudefibrecontent
loss in mass resulting from ashing of the dried residue obtained after acid and alkaline digestion of the sample by
theprocedure describedinthis InternationalStandard, divided bythe mass of thetest portion
NOTE The crudefibre content is expressed in gramsperkilogram. Itmayalsobeexpressedasa mass fraction in percent.
4P r i n c i p l e
The test portion is treated with boiling dilute sulfuric acid. The residue is separated by filtration, washed and thentreated with boiling potassium hydroxide solution. The residue is separated by filtration, washed, dried, weighedandthen ashed. Theloss inmass resulting from ashingcorresponds to the mass of crudefibrein the test portion.ISO 6865:2000(E)
2 © ISO2000 –All rights reserved5 Reagents andmaterials
Useonlyreagents of recognized analyticalgrade.
5.1 Water, complying with at least grade3 inaccordance with ISO3696.
5.2 Hydrochloric acid, c(HCl)=0,5mol/l.
5.3 Sulfuric acid, c(H2SO4)=(0 ,1 3 /Gb10,005)mol/l.
5.4 Potassium hydroxidesolution, c(KOH)=(0,23 /Gb10,005)mol/l.
5.5 Acetone.
5.6 Filteraid: seasand , or Celite ®1)545, or material of anequivalent quality.
Before use, treat sea sand with boiling hydrochloric acid [ c(HCl)=4mol/l], wash with water until free from acid and
heat at a temperatureof (500 /Gb125)°C for at least 1h.
Beforeuse, heat other filter aids at a temperatureof (500 /Gb125)°C for at least 4h.
5.7 Antifoam agent, for instance n-octanol.
5.8 Light petroleum, boilingrange 40 °Cto60°C.
6 Apparatus
Usual laboratoryapparatus and, in particular, thefollowing.
6.1 Grinding device, capable of grinding the sample so that it passes completely through a sievewith 1mm
apertures.
6.2 Analyticalbalance, withaccuracyof atleast0,1mg.
6.3 Filter crucibles, of quartz, porcelain or hard glass, with fused sintered filter plate with a pore size of 40 /G6dm
to100 /G6dm (porositygrade P100according toISO4793:1980[1]).
Before using for the first time, carefully and gradually heat a new filter crucible to a temperature not exceeding
525°C andleavefor afewminutes at(500 /Gb125)°C.
Stainless-steel crucibles with stainless-steel sieve plates with an aperture size of 90 /G6dm of identical performance
characteristics maybe usedas analternative.
6.4 Porcelain sieveplates.
6.5 Incineration dishes.6.6 Beakers orconical flasks, of 500ml capacity, provided with a suitable cooling device, for example a
condensor or adish.
1) Celite ®is the tradename of a commercially available product. This information is given for the convenience of users of this International
Standard and does not constitute an endorsement by ISO of this product. Equivalent products may be used if they canbeshown to lead to
th esa mer e su lt s.ISO 6865:2000(E)
© ISO2000 –All rights reserved 36.7 Drying oven, electrically heated and ventilated, capable of being maintained at a temperature of
(130 /Gb12)°C.
6.8 Desiccator, containing blue silicagel as desiccant, provided with a perforated plate, preferably of aluminium
or stainless steel, of thickness 2mm to3mm.
6.9 Muffle furnace, electrically heated, provided with air circulation and temperature control capable of
maintaining the temperaturearound the crucibles tothe nearest 25 °C at temperatures of 475 °Cto525°C.
The reading of the pyrometer of the muffle furnace cannot always be trusted: deviations may occur. Therefore, the
temperaturein the mufflefurnaceshould becheckedregularly.
Depending on the size and type of the muffle furnace, temperatures in the furnace may differ from one place to
another place. When the oven door is closed, adequate air supply should be guaranteed. The volume flow rate of
air should not beso great that material is carried awayfrom thecrucibles.
6.10 Cold-extraction device, provided with
/Gbeasupport for thefilter crucible(6.3);
/Gbeadischargepipe witha tap tothe vacuum andliquid outlet; and
/Gbeconnectingrings for connectingthefilter crucible(6.3).
6.11 Heating device (for manual method), provided with a suitable cooling device capable of maintaining the
volume constant during boiling.
6.12 Heating device (for the semi-automaticmethod) for acid and alkaline digestion, provided with
/Gbeasupport for thefilter crucible(6.3);
/Gbeadischargepipe witha tap tothe vacuum andliquid outlet;
/Gbeaboiling cylinder of at least 270mlcapacity, withareflux condenser;
/Gbeconnectingrings for connectingtheheating device tothefilter crucible(6.3) andtheboilingcylinder; and
/Gbeoptionally, provision for compressed air.
Beforeuse, preheat the apparatus with boiling water for 5min.
7 Sampling
Sampling is not part of the method specified in this International Standard. A recommended sampling method is
given in ISO 6497 [6].
It is important that the laboratory receive a sample which is truly representative and has not been damaged or
changedduringtransport or storage.
8 Preparationoftestsample
Prepare the test sampleinaccordance withISO6498.
Using the grinding device (6.1), grind the air-dry laboratory sample so that it passes completely through a sieve
with 1mm apertures. Mix thoroughly.ISO 6865:2000(E)
4 © ISO2000 –All rights reservedFor themanualmethod, proceed in accordance withclause9.
For thesemi-automatic method, proceedinaccordance withclause 10.
9 Procedurefor manualmethod
9.1 Test portion
Weighabout 1 gof the prepared test sample(clause 8) to the nearest 0,1mg( m1).
If the fat content of the sample exceeds 100g/kg, or if the sample contains fats which cannot be extracted directly
with light petroleum (5.8), transfer thesampletoacrucible(6.3) and proceedinaccordance with 9.2.
If the fat content of the sample does not exceed 100g/kg, transfer the sample to a beaker (6.6) and proceed in
accordance with 9.3 if the carbonate content, expressed as calcium carbonate, exceeds 50g/kg. If this is not thecase,proceedinaccordance with9.4.
9.2 Preliminarydefatting
In the cold-extraction device (6.10), defat the sample three times under vacuum with 30ml of light petroleum (5.8)
eachtime. Drytheresiduebysuction after each washing. Transfer theresiduetoabeaker (6.6).
If the carbonate content, expressed as calcium carbonate, exceeds 50g/kg, proceed in accordance with 9.3. If not,
proceed in accordance with9.4.
9.3 Removal of carbonate
Pour 100ml of hydrochloric acid (5.2) over the sample. Stir continuouslyfor 5min. Carefully pour the mixture into afilter crucible, the bottom of which is covered with a thin layer of filter aid (5.6).
Decant twice with 100mlof water eachtime. Takecaresothat as little materialas possible ends uponthefilter.Transfer thecontents of thecrucible tothe original beaker and proceedinaccordancewith9.4.
9.4 Acid digestion
Pour 150ml of sulfuric acid (5.3) over the sample.
Bring to the boil as quicklyas possible and continue steadyboiling for (30 /Gb11) min.
At the beginning of boiling, swirl a few times. If foaming occurs, add a few drops of antifoam agent (5.7). During
boiling, maintain a constant volume byusing a suitable cooling device (see 6.6 and 6.11).
9.5 Firstfiltration
In the filter crucible (6.3) apply a layer of filter aid (5.6) with a thickness of about one-fifth of the height of the filtercrucible. Thefilter aidmaybecovered byasieveplate (6.4) toprevent splashing.
When the boiling time has elapsed, filter the liquid down a stirring rod into the filter crucible. Apply a weak vacuum,
so that just in one pass 150ml is almost completely poured. If the filter blocks, carefully shift aside with a stirring
rodthe crudefibre coveringthefilter aid.
Wash the residue five times with about 10ml of hot water each time. Take care that the filter plate of the crucible
remains covered bythefilter aid, sothat thecrudefibre will not reachthefilter plate.ISO 6865:2000(E)
© ISO2000 –All rights reserved 5Release the vacuum and transfer a volume of acetone (5.5) which is sufficient to just cover the residue. Wait a few
minutes and then remove the acetone by applying slight suction. For a few moments suck air through to dry theresidue.
If thesamplecontains fats which cannot be extracted directly with light petroleum (5.8), proceed in accordance with
9.6. If not, proceed inaccordance with 9.7.
9.6 Defatting
In the cold-extraction device (6.10), defat the sample three times under vacuum with 30ml of light petroleum (5.8)eachtime. Drytheresiduebysuction after each washing.
9.7 Alkaline digestion
Transfer theresiduequantitativelyintothesamebeaker usedfor the acid digestion.
Add 150ml of potassium hydroxide solution (5.4) and bring to the boil as quickly as possible. Continue steady
boiling for (30 /Gb11)min. During boiling, maintain a constant volume by using a suitable cooling device (see 6.6 and
6.11).
9.8 Second filtration
Filter the contents of the beaker through the filter crucible (6.3) containing a layer of filter aid (5.6) with a thickness
of about one-fifthof the height of thefilter crucible, coveredbya sieve plate(6.4) to prevent splashing.
Washtheresidue withhot water untiltherinsings areneutral.
Wash the residue three times under vacuum with 30ml of acetone (5.5) each time. Dry the residue by suction after
each washing.
9.9 Drying
Place the filter crucible in an incineration dish (6.5) and dry the dish with its contents for at least 2h in the drying
oven(6.7) set at atemperature of 130 °C.
During ashing or cooling, parts of the sintered filter plate of the crucible may come loose. As this may cause an
incorrect analysis result, place thefilter crucible in an incineration dish.
Leave the filter crucible and the incineration dish to cool in the desiccator (6.8). Immediately after removal from the
desiccator, weighthefilter crucibleandthe incineration dish tothe nearest 0,1mg( m2).
9.10 Ashing
Place the filter crucible and the incineration dish in the muffle furnace (6.9) and ash its contents at a temperature of
(500 /Gb125)°C untilthedifferencebetweentwoconsecutive weighings after cooling does not exceed2mg.
After each ashing, leave the filter crucible and the incineration dish to cool partly and, while still warm, place in the
desiccator. Leave tocoolcompletelythen weighto thenearest 0,1mg( m3).
9.11 Blank determination
Carry out a blank determination as described in 9.4 to 9.10 with about the same quantity of filter aid (5.6) but
without thesample.
Theloss inmass resulting from ashing(9.10) shall not exceed 2mg.
Proceed in accordance with clause 11.ISO 6865:2000(E)
6 © ISO2000 –All rights reserved10 Procedurefor semi-automatic method
10.1 Test portion
Weigh about 1 g of the prepared test sample (clause 8) to the nearest 0,1mg ( m1). Transfer this quantity to a filter
crucible (6.3) provided with about 2g of filter aid (5.6).
If the fat content of the sample exceeds 100g/kg, or if the sample contains fats which cannot be extracted directly
with light petroleum (5.8), proceed in accordance with 10.2.
If the fat content of the sample does not exceed 100g/kg, proceed in accordance with 10.3 if the carbonate
content, expressed as calcium carbonate, exceeds 50g/kg. If this is not thecase, proceed in accordance with10.4.
10.2 Preliminarydefatting
Connect the filter crucible to the cold-extraction device (6.10) and wash the sample three times under vacuum with
30mlof light petroleum (5.8) eachtime. Drytheresidue bysuctionafter each washing.
If the carbonate content, expressed as calcium carbonate, exceeds 50g/kg, proceed in accordance with 10.3. If
not, proceed in accordance with10.4.
10.3 Removal of carbonate
Connect the filter crucible to the heating device (6.12). Wash the sample three times with 30ml of hydrochloric acid(5.2) each time. After eachaddition, leavefor about 1minbeforefiltering.
Washoncewith about 30mlof water andproceed inaccordance with 10.4.
10.4 Acid digestion
Connect the boiling cylinder to the filter crucible. Transfer 150 ml of boiling sulfuric acid (5.3) to the cylinder with the
filter crucible. If foaming occurs, add a few drops of antifoam agent (5.7). Bring to the boil as quickly as possible
and boil vigorouslyfor (30±1)min.
10.5 Firstfiltration
Switch off the heating. Open the tap to the discharge pipe and, under vacuum, filter the sulfuric acid through thefilter crucible. Wash the residue at least three times with about 30ml of hot water each time, until neutral. Dry theresidue bysuctionafter each washing.
If filtration problems occur, it is recommended to carefullyblow air through to remove blockage of the filter.If thesamplecontains fats which cannot be extracted directly with light petroleum (5.8), proceed in accordance with
10.6. If not, proceed in accordance with10.7.
10.6 Defatting
Connect the filter crucible to the cold-extraction device (6.10) and wash the residue three times under vacuum with
30ml of acetone (5.5) each time. Next, wash the residue three times under vacuum with 30ml of light petroleum(5.8) each time. Drytheresidue bysuctionafter each washing.
10.7 Alkaline digestion
Close the outlet tap. Transfer 150ml of boiling potassium hydroxide solution (5.4) to the cylinder with the filtercrucible. Add a few drops of antifoam agent (5.7). Bring to the boil as quickly as possible and boil vigorously for
(30/Gb11)min.ISO 6865:2000(E)
© ISO2000 –All rights reserved 710.8 Second filtration
Switch off the heating. Open the tap to the discharge pipe and under vacuum filter the potassium hydroxide
solution through the filter crucible. Wash the residue at least three times with about 30ml of hot water each time,untilneutral. Drytheresidue bysuctionafter each washing.
If filtration problems occur, it is recommended to carefullyblow air through to remove blockage of the filter.Connect the filter crucible to the cold-extraction device (6.10) and wash the sample three times under vacuum with
30mlof acetone(5.5) each time. Drytheresidue bysuctionafter each washing.
10.9 Drying
Place the filter crucible in an incineration dish (6.5) and dry the dish with its contents for at least 2h in the drying
oven(6.7) set at atemperature of 130 °C.
During ashing or cooling, parts of the sintered filter plate of the crucible may come loose. As this may cause an
incorrect analysis result, place thefilter crucible in an incineration dish.
Leave the filter crucible and the incineration dish to cool in the desiccator (6.8). Immediately after removal from the
desiccator, weighthefilter crucibleandthe incineration dish tothe nearest 0,1mg( m2).
10.10 Ashing
Place the filter crucible and the incineration dish in the muffle furnace (6.9) and ash its contents at a temperature of
(500/Gb125)°C untilthedifferencebetweentwoconsecutive weighings after cooling does not exceed2mg.
After each ashing, leave the filter crucible and the incineration dish to cool partly and, while still warm, place in the
desiccator. Leave tocoolcompletelythen weighto thenearest 0,1mg( m3).
10.11 Blank determination
Carry out a blank determination as described in 10.4 to 10.10 with about the same quantity of filter aid (5.6) but
without thesample.
Theloss inmass resulting from ashing(10.10) shallnot exceed2mg.
11 Calculation
Calculate the crudefibre content of thetest samplebythe equation:
23
f
1mmwm/G2d/G3d
where
wfis thecrudefibrecontent, ingrams per kilogram, of thetest sample;
m1is themass, ingrams, of thetest portion (9.1 or 10.1);
m2is the mass, inmilligrams, of the incineration dish with the filter crucible with the residue obtained after
dryingat 130 °C (9.9 or 10.9);
m3is the mass, inmilligrams, of the incineration dish with the filter crucible with the residue obtained after
ashingat (500 /Gb125)°C (9.10 or 10.10).ISO 6865:2000(E)
8 © ISO2000 –All rights reservedRoundtheresultto the nearest1g/kg.
NOTE The resultmayalso becalculated as amass fractionin percent.
12 Precision
12.1 Interlaboratorytests
Details of interlaboratory tests on the precision of the method are given in annexA. The values derived from these
tests maynot be applicabletoconcentrationranges andmatrices other thanthosegiven.
12.2 Repeatability
The absolute difference between two independent single test results, obtained using the same method on identical
test material in the same laboratory by the same operator using the same equipment within a short interval of time,
willinnot morethan 5% of cases exceedtherepeatabilitylimit ( r) mentioned in or derivedfrom Table1.
Table1—Repeatabilitylimit (r)and reproducibilitylimit (R)
Sample Crudefibrecontent rR
g/kg g/kg g/kg
Sunflower seed meal 223,3 8,4 16,1
Palm kernelexpellers 190,3 19,4 42,5
Cattlefeed(pelleted) 115,8 5,3 13,8
Cornglutenfeed 73,3 5,8 9,1
Tapioca 60,2 5,6 8,8
Dogfood 30,0 3,2 8,9
Catfood 22,8 2,7 6,4
12.3 Reproducibility
The absolute difference between two single test results, obtained using the same method on identical test material
in different laboratories by different operators using different equipment, will in not more than 5% of cases exceed
the reproducibilitylimit ( R) mentioned in or derivedfrom Table1.
13 Testreport
Thetest report shallspecify:
/Gbeall information necessaryfor the complete identification of the sample;
/Gbethesamplingmethodusedif known;
/Gbethetest methodused, withreferencetothis International Standard;
/Gbeall operating details not specified in this International Standard, or regarded as optional, together with details of
anyincidents whichmayhave influenced the test result(s);
/Gbethetest result obtained, or thetwo test results obtained if therepeatabilityhas beenchecked.ISO 6865:2000(E)
© ISO2000 –All rights reserved 9Annex A
(informative)
Results ofinterlaboratorytests
The precision of the method was established in 1996 and 1997 by four interlaboratory tests carried out in
accordance with the split-level experiment of ISO5725:1986 [2]2)with the exception of application of the Grubbs
outliertestinstead of theDixon outliertest.TheGrubbsoutliertest isdescribedinISO5725-2:1994[4].
In the interlaboratory tests 10 laboratories participated. Samples of sunflower seed meal, palm kernel expellers,
cattlefeed(pelleted), cornglutenfeed, tapioca, dogfoodandcat food wereinvestigated.
TableA.1 —Statistical resultsof interlaboratorytests
Parameter Samplea
1234567
Number of laboratories retained after eliminating
outliers1 091 01 01 01 01 0
Mean crudefibre content, g/kg 223,3 190,3 115,8 73,3 60,2 30,0 22,8
Repeatabilitystandard deviation( sr), g/kg 3,00 6,93 1,89 2,07 2,00 1,14 0,96
Repeatabilitycoefficient of variation, % 1,3 3,6 1,6 2,8 3,3 3,8 4,2
Repeatabilitylimit ( r)[r=2 , 8/Gb4sr),g/kg 8,4 19,4 5,3 5,8 5,6 3,2 2,7
Reproducibilitystandard deviation ( sR), g/kg 5,75 15,18 4,93 3,25 3,14 3,18 2,29
Reproducibilitycoefficient of variation, % 2,6 8,0 4,3 4,4 5,2 10,6 10,0
Reproducibilitylimit ( R)[R=2,8/Gb4sR), g/kg 16,1 42,5 13,8 9,1 8,8 8,9 6,4
a1: sunflower seedmeal;
2: palm kernelexpellers;
3: cattlefeed(pelleted);
4: cornglutenfeed;5: tapioca;6: dogfood;7: cat food.
2) ISO5725:1986 has nowbeen withdrawn.ISO 6865:2000(E)
10 © ISO2000 –All rights reservedBibliography
[1] ISO 4793:1980, Laboratory sintered(fritted) filters —Porosity grading, classificationand designation .
[2] ISO 5725:1986, Precision of test methods —Determination of repeatability and reproducibility for a
standard testmethodby inter-laboratory tests .
[3] ISO 5725-1:1994, Accuracy (trueness and precision) of measurement methods and results —Part 1:
General principles and definitions .
[4] ISO 5725-2:1994, Accuracy (trueness and precision) of measurement methods and results —Part 2: Basic
metodfor the determination of repeatability andreproducibility of astandardmeasurement method.
[5] ISO 6492, Animalfeedingstuffs —Determination of fat content .
[6] ISO 6497, Animalfeedingstuffs —Sampling .
[7] ISO 6541, Agricultural food products —Determination of crude fibre content —Modified Scharrer method .ISO 6865:2000(E)
ICS 65.120
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© ISO2000 –All rights reserved