First Commission Directive

++++

of 13 November 1979

KOMMISSIONENS FOERSTE DIREKTIV

laying down Community methods of analysis for testing certain partly or wholly dehydrated preserved milk for human consumption

af 13 . november 1979

(79/1067/EEC)

om fastlaeggelse af faellesskabsanalysemetoder til kontrol af visse former for konserveret helt eller delvist inddampet maelk bestemt til konsum

THE COMMISSION OF THE EUROPEAN COMMUNITIES,

( 79/1067/EOEF )

Having regard to the Treaty establishing the European Economic Community,

KOMMISSIONEN FOR DE EUROPAEISKE FAELLESSKABER HAR -

Having regard to Council Directive 76/118/EEC of 18 December 1975 on the approximation of the laws of the Member States relating to certain partly or wholly dehydrated preserved milk for human consumption [1], and in particular Article 11 thereof,

under henvisning til traktaten om oprettelse af Det europaeiske oekonomiske Faellesskab ,

Whereas under Article 11 of Directive 76/118/EEC, the composition of certain partly or wholly dehydrated preserved milk is required to be verified by Community methods of analysis;

under henvisning til Raadets direktiv 76/118/EOEF af 18 . december 1975 om indbyrdes tilnaermelse af medlemsstaternes lovgivning vedroerende visse former for konserveret helt eller delvist inddampet maelk bestemt til menneskefoede ( 1 ) , saerlig artikel 11 , og

Whereas it is desirable to adopt an initial series of methods in respect of which studies have been completed;

ud fra foelgende betragtninger :

Whereas the measures provided for in this Directive are in accordance with the opinion of the Standing Committee of Foodstuffs,

I henhold til artikel 11 i direktiv 76/118/EOEF skal sammensaetningen af visse former for konserveret helt eller delvist inddampet maelk kontrolleres efter faellesskabsanalysemetoder ;

HAS ADOPTED THIS DIRECTIVE:

en foerste raekke af metoder , for hvilke undersoegelserne er sluttet , boer vedtages ;

Article 1

de i dette direktiv fastsatte foranstaltninger er i overensstemmelse med udtalelse fra Den staaende Levnedsmiddelkomité -

Member States shall take all measures necessary to ensure that the analyses necessary for verification of the criteria set out in Annex I are carried out in accordance with the methods described in Annex II.

UDSTEDT FOELGENDE DIREKTIV :

Article 2

Artikel 1

Where alternative methods for a single determination are specified, the sample may be analysed by either method. The test report referred to in Annex II must state the method which has been employed.

Medlemsstaterne foreskriver , at de fornoedne analyser til efterproevning af de i bilag I angivne egenskaber , udfoeres efter de i bilag II beskrevne metoder .

Article 3

Artikel 2

Member States shall bring into force the laws, regulations and administrative provisions necessary to comply with this Directive within 18 months of its notification. They shall forthwith inform the Commission thereof.

Naar der er anfoert alternative metoder for en enkelt bestemmelse , kan analysen foretages efter hvilken metode , der oenskes . Den i bilag II omhandlede afproevningsrapport skal angive , hvilken metode der er anvendt .

Article 4

Artikel 3

This Directive is addressed to the Member States.

Medlemsstaterne saetter de noedvendige administrativt eller ved lov fastsatte bestemmelser i kraft for senest 18 maaneder efter dets meddelelse at efterkomme dette direktiv . De underretter straks Kommissionen herom .

Artikel 4

Done at Brussels, 13 November 1979.

Dette direktiv er rettet til medlemsstaterne .

For the Commission

Udfaerdiget i Bruxelles , den 13 . november 1979 .

Étienne Davignon

Paa Kommissionens vegne

Member of the Commission

Etienne DAVIGNON

[1] OJ No L 24, 30. 1. 1976, p. 49.

Medlem af Kommissionen

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( 1 ) EFT nr . L 24 af 30 . 1 . 1976 , s . 49 .

ANNEX I

BILAG I

SCOPE OF THE FIRST COMMUNITY METHODS OF ANALYSIS FOR CERTAIN PARTLY OR . WHOLLY DEHYDRATED PRESERVED MILK DIRECTIVE

ANVENDELSESOMRAADET FOR DE FOERSTE FAELLESSKABSANALYSEMETODER SOM ANVENDES I DIREKTIVET FOR VISSE FORMER FOR KONSERVERET HELT ELLER DELVIS INDDAMPET MAELK

I. General provisions

I . Almindelige bestemmelser

II. Determination of dry matter in:

II . Bestemmelse af toerstof i :

- unsweetened condensed high fat milk (using method 1, Annex II),

- kondenseret maelk med hoejt fedtindhold eller usukret kondenseret maelk med hoejt fedtindhold ( under anvendelse af metode 1 , bilag II ) ,

- unsweetened condensed milk (using method 1, Annex II),

- kondenseret maelk , usukret kondenseret maelk eller kondenseret soedmaelk ( under anvendelse af metode 1 , bilag II ) ,

- unsweetened condensed partly skimmed milk (using method 1, Annex II),

- kondenseret delvis skummet maelk eller usukret kondenseret delvis skummet maelk ( under anvendelse af metode 1 , bilag II ) ,

- unsweetened condensed skimmed milk (using method 1, Annex II),

- kondenseret skummetmaelk eller usukret kondenseret skummetmaelk ( under anvendelse af metode 1 , bilag II ) ,

- sweetened condensed milk (using method 1, Annex II),

- sukret kondenseret maelk eller sukret kondenseret soedmaelk ( under anvendelse af metode 1 , bilag II ) ,

- sweetened condensed partly skimmed milk (using method 1, Annex II),

- sukret kondenseret delvis skummet maelk ( under anvendelse af metode 1 , bilag II ) ,

- sweetened condensed skimmed milk (using method 1, Annex II).

- sukret kondenseret skummetmaelk ( under anvendelse af metode 1 , bilag II ) .

III. Determination of moisture in:

III . Bestemmelse af vandindhold i :

- dried high fat milk or high fat milk powder (using method 2, Annex II), — dried whole milk or whole milk powder (using method 2, Annex II),

- maelkepulver med hoejt fedtindhold ( under anvendelse af metode 2 , bilag II ) ,

- dried partly skimmed milk or partly skimmed-milk powder (using method 2, Annex II),

- maelkepulver eller soedmaelkspulver ( under anvendelse af metode 2 , bilag II ) ,

- dried skimmed milk or skimmed-milk powder (using method 2, Annex II).

- pulver af delvis skummet maelk ( under anvendelse af metode 2 , bilag II ) ,

IV. Determination of fat in:

- skummetmaelkspulver ( under anvendelse af metode 2 , bilag II ) .

- unsweetened condensed high fat milk (using method 3, Annex II),

IV . Bestemmelse af fedt i :

- unsweetened condensed milk (using method 3, Annex II),

- kondenseret maelk med hoejt fedtindhold eller usukret kondenseret maelk med hoejt fedtindhold ( under anvendelse af metode 3 , bilag II ) ,

- unsweetened condensed partly skimmed milk (using method 3, Annex II),

- kondenseret maelk , usukret kondenseret maelk eller kondenseret soedmaelk ( under anvendelse af metode 3 , bilag II ) ,

- unsweetened condensed skimmed milk (using method 3, Annex II),

- kondenseret delvis skummet maelk eller usukret kondenseret delvis skummet maelk ( under anvendelse af metode 3 , bilag II ) ,

- sweetened condensed milk (using method 3, Annex II),

- kondenseret skummetmaelk eller usukret kondenseret skummetmaelk ( under anvendelse af metode 3 , bilag II ) ,

- sweetened condensed partly skimmed milk (using method 3, Annex II),

- sukret kondenseret maelk eller sukret kondenseret soedmaelk ( under anvendelse af metode 3 , bilag II ) ,

- sweetened condensed skimmed milk (using method 3, Annex II),

- sukret kondenseret delvis skummet maelk ( under anvendelse af metode 3 , bilag II ) ,

- dried high fat milk or high fat milk powder (using method 4, Annex II),

- sukret kondenseret skummetmaelk ( under anvendelse af metode 3 , bilag II ) ,

- dried whole milk or whole milk powder (using method 4, Annex II),

- maelkepulver med hoejt fedtindhold ( under anvendelse af metode 4 , bilag II ) ,

- dried partly skimmed milk or partly skimmed-milk powder (using method 4, Annex II),

- maelkepulver eller soedmaelkspulver ( under anvendelse af metode 4 , bilag II ) ,

- dried skimmed milk or skimmed-milk powder (using method 4, Annex II).

- pulver af delvist skummet maelk ( under anvendelse af metode 4 , bilag II ) ,

V. Determination of sucrose in:

- skummetmaelkspulver ( under anvendelse af metode 4 , bilag II ) .

- sweetened condensed milk (using method 5, Annex II),

V . Bestemmelse af saccharose i :

- sweetened condensed partly skimmed milk (using method 5, Annex II),

- sukret kondenseret maelk eller sukret kondenseret soedmaelk ( under anvendelse af metode 5 , bilag II ) ,

- sweetened condensed skimmed milk (using method 5, Annex II).

- sukret kondenseret delvis skummet maelk ( under anvendelse af metode 5 , bilag II ) ,

VI. Determination of lactic acid and lactates in:

- sukret kondenseret skummetmaelk ( under anvendelse af metode 5 , bilag II ) .

- dried high fat milk or high fat milk powder (using method 6, Annex II),

VI . Bestemmelse af maelkesyre og laktater i :

- dried whole milk or whole milk powder (using method 6, Annex II),

- maelkepulver med hoejt fedtindhold ( under anvendelse af metode 6 , bilag II ) ,

- dried partly skimmed milk or partly skimmed-milk powder (using method 6, Annex II),

- maelkepulver eller soedmaelkspulver ( under anvendelse af metode 6 , bilag II ) ,

- dried skimmed milk or skimmed-milk powder (using method 6, Annex II).

- pulver af delvis skummet maelk ( under anvendelse af metode 6 , bilag II ) ,

VII. Determination of phosphatase activity in:

- skummetmaelkspulver ( under anvendelse af metode 6 , bilag II ) .

- dried high fat milk or high fat milk powder (using method 7 or 8, Annex II),

VII . Bestemmelse af phosphataseaktivitet i :

- dried whole milk or whole milk powder (using method 7 or 8, Annex II),

- maelkepulver med hoejt fedtindhold ( under anvendelse af metode 7 eller 8 , bilag II ) ,

- dried partly skimmed milk or partly skimmed-milk powder (using method 7 or 8, Annex II),

- maelkepulver eller soedmaelkspulver ( under anvendelse af metode 7 eller 8 , bilag II ) ,

- dried skimmed milk or skimmed-milk powder (using method 7 or 8, Annex II).

- pulver af delvis skummet maelk ( under anvendelse af metode 7 eller 8 , bilag II ) ,

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- skummetmaelkspulver ( under anvendelse af metode 7 eller 8 , bilag II ) .

ANNEX II

BILAG II

METHODS OF ANALYSIS RELATING TO THE COMPOSITION OF CERTAIN PARTLY OR WHOLLY DEHYDRATED PRESERVED MILK PRODUCTS INTENDED FOR HUMAN CONSUMPTION

ANALYSEMETODER I FORBINDELSE MED FORBRUGET AF VISSE FORMER FOR KONSERVERET HELT ELLER DELVIS INDDAMPET MAELK BESTEMT TIL KONSUM

GENERAL PROVISIONS

ALMINDELIGE BESTEMMELSER

1. PREPARATION OF THE SAMPLE FOR CHEMICAL ANALYSIS

1 . FREMSTILLING AF PROEVE TIL KEMISKE ANALYSER

1.1. Unsweetened condensed high fat milk

1.1 . Kondenseret maelk med hoejt fedtindhold eller usukret kondenseret maelk med hoejt fedtindhold

Unsweetened condensed milk

Kondenseret maelk , usukret kondenseret maelk eller kondenseret soedmaelk

Unsweetened condensed partly skimmed milk

Kondenseret delvis skummet maelk eller usukret kondenseret delvis skummet maelk

Unsweetened condensed skimmed milk

Kondenseret skummetmaelk eller usukret kondenseret skummetmaelk

Shake and invert the closed can. Open the can and slowly pour the milk into a second container which can be closed hermetically, mixing by repeated transfer. Ensure that all remaining fat and milk adhering to the wall and the ends of the can are mixed in with the sample. Close the container. If the contents are not homogeneous warm the container in a waterbath at 40 oC. Shake vigorously every 15 minutes. After two hours, remove the container from the water-bath and let it cool to room temperature. Remove the lid and thoroughly mix the contents of the container with a spoon or spatula (if the fat has separated the sample should not be tested). Store in a cool place.

Den lukkede daase omrystes og vendes . Daasen aabnes og maelken haeldes langsomt op i en anden beholder , der kan lukkes hermetisk , og der blandes ved gentagne omhaeldninger . Det sikrer , at tilbagebleven maelk og fedt , der sidder fast paa daasens side og ender , blandes op i proeven . Beholderen lukkes . Hvis indholdet ikke er homogent , varmes beholderen paa et vandbad ved 40 * . Der omrystes omhyggeligt hvert 15 . min . Efter 2 timer fjernes beholderen fra vandbadet , og den afkoeles til stuetemperatur . Laaget fjernes og der blandes omhyggeligt med en ske eller en spatel ( hvis fedtet er udskilt , kan proeven ikke analyseres ) . Proeven opbevares koeligt .

1.2. Sweetened condensed milk

1.2 . Sukret kondenseret maelk eller sukret kondenseret soedmaelk

Sweetened condensed partly skimmed milk

Sukret kondenseret delvis skummet maelk

Sweetened condensed skimmed milk

Sukret kondenseret skummetmaelk

Cans: Warm the closed can in a waterbath at 30 to 40 oC for about 30 minutes. Open the can and thoroughly mix the contents with a spatula or a spoon by making upward, downward, and circular movements in order to obtain an intimate mixture of the top and bottom layers with all the contents. Ensure that the remaining milk adhering to the wall and end of the can is incorporated in the sample. As far as possible, pour the contents into a second container provided with an air-tight lid. Close the container and store in a cool place.

Daaser : Den lukkede daase opvarmes paa et vandbad ved 30 - 40 * C i ca . 30 min . Daasen aabnes og indholdet blandes omhyggeligt med en spatel eller en ske , idet der foretages bevaegelser op og ned og rundt for at opnaa en god opblanding af de oevre og nedre lag med det oevrige indhold . Det sikres , at den tilbageblevne maelk , der sidder fast paa daasens vaegge og ender er blandet op i proeven . Om muligt haeldes indholdet over i en anden beholder , der er udstyret med et lufttaet laag . Beholderen lukkes og opbevares koeligt .

Tubes: Cut the end and pour the contents into a container provided with an air-tight lid. Next, cut the tube lengthwise. Scrape out all material adhering to the interior and mix it carefully with the rest of the contents. Store the container in a cool place.

Tuber : Enden skaeres af og indholdet haeldes over i en anden beholder , der er udstyret med et lufttaet laag . Tuben skaeres dernaest op paa langs . Al materiale , der sidder fast indvendigt skrabes af og blandes omhyggeligt op i resten af indholdet . Beholderen opbevares koeligt .

1.3. Dried high fat milk or high fat milk powder

1.3 . Maelkepulver med hoejt fedtindhold

Dried whole milk or whole milk powder

Maelkepulver eller soedmaelkspulver

Dried partly skimmed milk or partly skimmed-milk powder

Pulver af delvis skummet maelk

Dried skimmed milk or skimmed-milk powder

Skummetmaelkspulver

Transfer the milk powder to a clean, dry container (with air-tight lid) of a capacity of twice the volume of the powder. Close the container immediately and thoroughly mix the milk powder by repeatedly shaking and inverting the container. During the preparation of the sample exposure of the milk powder to the atmosphere should be avoided as far as possible to minimize absorption of moisture.

Maelkepulveret overfoeres til en ren , toer beholder ( med lufttaet laag ) med et rumfang paa to gange rumfanget af pulveret . Beholderen lukkes straks , og maelkepulveret blandes omhyggeligt , idet beholderen gentagne gange rystes og vendes . Under fremstillingen af proeven maa maelkepulver ikke bringes i kontakt med atmosfaeren mere end hoejst noedvendigt for at minimere optagelsen af fugt .

2. REAGENTS

2 . REAGENSER

2.1. Water

2.1 . Vand

2.1.1. Wherever mention is made of water for dissolution, dilution or washing purposes, distilled water, or demineralized water of at least equivalent purity shall be used.

2.1.1 . Hvor der naevnes vand til oploesning , fortynding eller udvaskning , skal der anvendes destilleret vand eller demineraliseret vand af mindst samme kvalitet .

2.1.2. Wherever reference is made to "dissolution", "solution" or "dilution" without further indication, "dissolution in water", "solution in water" and "dilution in water" is meant.

2.1.2 . Hvor der refereres til " oploesning " , " en oploesning " eller " fortynding " uden yderligere angivelse , menes der " oploesning i vand " , " en vandig oploesning " og " fortynding med vand " .

2.2. Chemicals

2.2 . Kemikalier

All chemicals used shall be of recognized analytical reagent quality except where otherwise specified.

Alle kemikalier skal vaere af anerkendt analysekvalitet , medmindre andet er specificeret .

3. EQUIPMENT

3 . APPARATUR

3.1. Lists of equipment

3.1 . Udstyrsliste

The lists of equipment contain only those items with a specialized use and items with a particular specification.

Udstyrslisterne indeholder kun de ting , der anvendes til noget specielt , og de ting , der skal vaere af en speciel kvalitet .

3.2. Analytical balance

3.2 . Analysevaegt

Analytical balance means a balance capable of weighing to at least 0,1 mg.

Ved en analysevaegt forstaas en vaegt , der kan veje med en noejagtighed paa 0,1 mg .

4. EXPRESSION OF RESULTS

4 . PRAESENTATION AF RESULTATERNE

4.1. Calculation of percentage

4.1 . Beregning af procentindhold

Except where otherwise specified, the results shall be calculated as a percentage by mass of the sample as received by the laboratory.

Medmindre andet er anfoert , skal resultaterne beregnes som en procentdel paa vaegtbasis af proeven , som den er modtaget af laboratoriet .

4.2. Number of significant figures

4.2 . Antal betyden cifre

The result shall not contain more significant figures than are justified by the precision of the method of analysis used.

Resultatet skal ikke indeholde flere betydende cifre , end det kan retfaerdiggoeres ud fra noejagtigheden af den anvendte analysemetode .

5. TEST REPORT

5 . AFPROEVNINGSRAPPORT

The test report shall identify the method of analysis used as well as the results obtained. In addition, it shall mention all details of procedure not specified in the method of analysis, or which are optional, as well as any circumstances that may have influenced the results obtained.

Afproevningsrapporten skal angive den anvendte analysemetode samt de opnaaede resultater . Derudover skal den anfoere alle de detaljer i fremgangsmaaden , der ikke er specificeret i analysemetoden , eller som er valgfrie , samt de omstaendigheder , der kan have oevet indflydelse paa det opnaaede resultat .

The test report shall give all the information necessary for the complete identification of the sample.

Afproevningsrapporten skal indeholde alle de oplysninger , der er noedvendige til en fuldstaendig identifikation af proeven .

METHOD 1: DETERMINATION OF DRY MATTER CONTENT

METODE 1 : BESTEMMELSE AF TOERSTOFINDHOLD

(oven 99 oC)

( ovn ved 99 * C )

1. SCOPE AND FIELD OF APPLICATION

1 . OMFANG OG ANVENDELSESOMRAADE

This method determines the dry matter content of:

Denne metode anvendes ved bestemmelse af toerstofindhold i :

- unsweetened condensed high fat milk,

- kondenseret maelk med hoejt fedtindhold eller usukret kondenseret maelk med hoejt fedtindhold ,

- unsweetened condensed milk,

- kondenseret maelk , usukret kondenseret maelk eller kondenseret soedmaelk ,

- unsweetened condensed partly skimmed milk,

- kondenseret delvis skummet maelk eller usukret kondenseret delvis skummet maelk ,

- unsweetened condensed skimmed milk,

- kondenseret skummetmaelk eller usukret kondenseret skummetmaelk ,

- sweetened condensed milk,

- sukret kondenseret maelk eller sukret kondenseret soedmaelk ,

- sweetened condensed partly skimmed milk,

- sukret kondenseret delvis skummet maelk ,

- sweetened condensed skimmed milk.

- sukret kondenseret skummetmaelk .

2. DEFINITION

2 . DEFINITION

The dry matter content of condensed milks: dry matter content as determined by the method specified.

Toerstofindholdet i kondenseret maelk : toerstofindholdet som det bestemmes ved den specificerede metode .

3. PRINCIPLE

3 . PRINCIP

A known amount of the sample is diluted with water, mixed with sand and dried at a temperature of 99 oC ± 1 oC. The mass after drying is the mass of dry matter and is calculated as a percentage by mass of the sample.

En kendt maengde af proeven fortyndes med vand , blandes med sand og toerres ved en temperatur paa 99 mere aller mindre 1 * C . Vaegten efter toerringen er toerstofvaegten og beregnes som en vaegtprocentdel af proeven .

4. REAGENTS

4 . REAGENSER

Quartz sand or sea sand, treated with hydrochloric acid (size of the grains: 0,18 to 0,5 mm, that is passing through a 500 micron sieve and retained by a 180 micron sieve). It should meet the following control test:

Quartssand eller havsand , behandlet med saltsyre ( kornstoerrelse : 0,18 - 0,35 mm , det vil sige , at det skal passere gennem en sigte med 500 micronmasker og holdes tilbage af en sigte med 180 micronmasker ) . Det skal opfylde foelgende kontrolproeve :

Heat about 25 g of sand for two hours in the drying oven (5.3) as described in 6.1. to 6.3. Add 5 ml of water, heat again in the oven for two hours, cool and reweigh. The difference between the two masses should not exceed 0,5 mg.

Ca . 25 g sand opvarmes 2 timer i toerreovn ( 5.3 . ) som beskrevet under 6.1 . til 6.3 . Derefter tilsaettes 5 ml vand , og der opvarmes igen i ovnen i 2 timer . Efter afkoeling vejes og forskellen mellem de to vaegte maa ikke overstige 0,5 mg .

If necessary treat the sand with a 25 % hydrochloric acid solution for three days, mixing occasionally. Wash with water until the acid reaction disappears or the wash water is chloride free. Dry at 160 oC and re-test as above.

Sandet behandles om noedvendigt med en 25 % saltsyreoploesning i tre dage ; der blandes lejlighedsvis . Vask med vand , indtil der ikke laengere er syrereaktion , eller indtil vaskevandet er chloridfrit . Der toerres ved 160 * C og foretages ny afproevning som ovenfor .

5. APPARATUS

5 . APPARATUR

5.1. Analytical balance.

5.1 . Analysevaegt

5.2. Metal dishes, preferably of nickel, aluminium or stainless steel. The dishes must have lids which fit very well but which are readily removed. Suitable dimensions are: diameter 60 to 80 mm and depth about 25 mm.

5.2 . Metalskaale , fortrinsvis af nikkel , aluminium eller rustfrit staal . Skaalene skal have laag , der passer godt , men som let kan fjernes . Passende dimensioner er : diameter 60 - 80 mm og dybde ca . 25 mm .

5.3. Atmospheric-pressure drying oven, well ventilated, thermostatically controlled with temperature regulated at 99 oC ± 1 oC. The temperature should be uniform throughout the oven.

5.3 . Toerreovn ved atmosfaeretryk , godt ventileret , termostatreguleret med temperaturen reguleret til 99 mere aller mindre 1 * C . Temperaturen skal vaere den samme i hele ovnen .

5.4. Desiccator, containing freshly activated silica gel with a water content indicator or an equivalent desiccant.

5.4 . Ekssikator , indeholdende frisk aktiveret silica gel med en indikator for vandindhold eller et tilsvarende toerremiddel .

5.5. Glass rods, flattened at one end of such a length that they can fit inside the metal dishes (5.2).

5.5 . Glasstave , fladtrykte i den ene ende og af en saadan laengde , at de kan ligge inden i metalskaalene ( 5.2 . ) .

5.6. Waterbath, boiling.

5.6 . Vandbad , kogende .

6. PROCEDURE

6 . FREMGANGSMAADE

6.1. Place about 25 g sand (4) and a short glass rod (5.5) in the dish (5.2).

6.1 . Ca . 25 g sand ( 4 ) og en kort glasstav ( 5.5 . ) anbringes i skaalen ( 5.2 . ) .

6.2. Without covering the dish and contents with the lid, place the dish, contents and lid in the oven (5.3) and heat for two hours.

6.2 . Uden at daekke skaalen og indholdet med laaget , anbringes skaalen , indholdet og laaget i ovnen ( 5.3 . ) i 2 timer .

6.3. Replace lid and transfer the dish to the desiccator (5.4). Allow to cool to room temperature and accurately weigh to the nearest 0,1 mg (M0).

6.3 . Laaget paasaettes , og skaalen flyttes over i ekssikkatoren ( 5.4 . ) . Man lader den afkoele til stuetemperatur , og den vejes med en noejagtighed paa 0,1 mg ( M0 ) .

6.4. Tilt the sand to one side of the dish. Introduce into the clear space about 1,5 g of the sample of sweetened condensed milk and 3,0 g of unsweetened condensed milk. Replace the lid and accurately weigh to the nearest 0,1 mg (M1).

6.4 . Idet laaget vippes , rystes sandet over til den ene side i skaalen . Ca . 1,5 g af proeven fyldes ned i det tomme rum , hvis det er sukret kondenseret maelk , og ca . 3,0 g , hvis det er usukret kondenseret maelk . Laaget paasaetes igen , og der vejes med en noejagtighed paa 0,1 mg ( M1 ) .

6.5. Remove the lid, add 5 ml of water and, with the aid of the glass rod, mix the liquids and then the sand and the liquid portion. Leave the rod in the mixture.

6.5 . Laaget fjernes , 5 ml vand tilsaettes , hvorefter foerst vaesken og derpaa sandet og vaesken blandes ved hjaelp af glasstaven . Staven efterlades i blandingen .

6.6. Place the dish on a boiling waterbath (5.6) until the water has evaporated; this usually takes 20 minutes. Stir the mixture from time to time with the rod to keep the mass well aerated so that the mass when dry does not form a cake. Lay the rod inside the dish.

6.6 . Skaalen anbringes paa et kogende vandbad . ( 5.6 . ) , indtil vandet er fordampet ; dette tager saedvanligvis ca . 20 min . Roer fra tid til anden i blandingen med staven , saaledes at massen ikke danner en kage . Laeg staven inde i skaalen .

6.7. Place the dish and lid in the oven for one and a half hours.

6.7 . Anbring skaalen og laaget i ovnen i ca . 1 time og 30 min .

6.8. Replace the lid, transfer the dish to the desiccator (5.4), allow to cool to room temperature and accurately weigh to the nearest 0,1 mg.

6.8 . Laaget paasaettes igen , skaalen flyttes over i ekssikkatoren , man lader den afkoele til stuetemperatur og der vejes med en noejagtighed paa 0,1 mg .

6.9. Replace the dish and lid in the oven, uncover the dish and heat it with its lid for a further hour.

6.9 . Skaalen og laaget anbringes atter i ovnen , laaget tages af skaalen og den og laaget opvarmes i yderligere 1 time .

6.10. Repeat process 6.8.

6.10 . Procedure 6.8 . gentages .

6.11. Repeat the described processes 6.9 and 6.10 until the difference in mass of two successive weighings is less than 0.5 mg or until the mass increases. If an increase in mass occurs use the lowest mass obtained in the calculation (7.1). Let the final weight recorded be M2 g.

6.11 . Procedure 6.9 . og 6.10 . gentages indtil forskellen mellem to paa hinanden foelgende vejninger er mindre end 0,5 mg eller indtil vaegten stiger . Hvis der forekommer en stigning i vaegten , anvendes den laveste vaegt ( 7.1 . ) . Den sidste vaegt kaldes M2 g .

7. EXPRESSION OF RESULTS

7 . PRAESENTATION AF RESULTATERNE

7.1. Method of calculation

7.1 . Beregningsmetode

The content of dry matter, calculated as a percentage by mass of the sample, is given by:

Toerstofindholdet beregnet som en vaegtprocent af proeven er givet ved :

×100

( M2 - M0 ) / ( M1 - M0 ) gange 100

where:

hvor :

M0 = mass, in g of the dish, lid and sand after process 6.3;

M0 = vaegten i g af skaal , laag og sand efter procedure 6.3 . ;

M1 = mass, in g of the dish, lid, sand and sample after process 6.4;

M1 = vaegten i g af skaal , laag , sand og proeve efter procedure 6.4 . ;

M2 = mass, in g of the dish, lid, sand and dried sample after process 6.11.

M2 = vaegten i g af skaal , laag , sand og toerret proeve efter procedure 6.11 .

7.2. Repeatability

7.2 . Repetitionsevne

Forskellen mellem resultaterne for to bestemmelser udfoert samtidigt eller hurtigt efter hinanden af den samme analytiker , der anvender den samme proeve under de samme betingelser , maa ikke overstige 0,2 g toerstof pr . 100 g produkt .

8. CALCULATION OF TOTAL MILK SOLIDS AND MILK SOLIDS NOT FAT

8 . BEREGNING AF TOTALT MAELKETOERSTOF OG FEDTFRIT MAELKETOERSTOF

8.1. The total milk solids content of the sweetened condensed milk is given by:

8.1 . Totalt maelketoerstofindhold af sukret kondenseret maelk er givet ved :

Total dry matter (obtained by method 1, Annex II) — sucrose (obtained by method 5, Annex II).

Totalt toerstof ( opnaaet ifoelge metode 1 , bilag II ) - saccharose ( opnaaet ifoelge metode 5 , bilag II ) .

8.2. The milk solids not fat content of the sweetened condensed milks is given by:

8.2 . Fedtfrit maelketoerstofindhold af sukret kondenseret maelk er givet ved :

Total dry matter (obtained by method 1, Annex II) — (sucrose content obtained by method 5, Annex II) and fat content (obtained by method 3, Annex II).

Totalt toerstof ( opnaaet ifoelge metode 1 , bilag II ) - ( saccharoseindhold opnaaet ifoelge metode 5 , bilag II ) og fedtindhold ( opnaaet ifoelge metode 3 , bilag II ) .

8.3. The milk solids not fat content of unsweetened condensed milks is given by:

8.3 . Fedtfrit maelketoerstofindhold af usukret kondenseret maelk er givet ved :

Total dry matter (obtained by method 1, Annex II) — fat content (obtained by method 3, Annex II).

Totalt toerstof ( opnaaet ifoelge metode 1 , bilag II ) - fedtindhold ( opnaaet ifoelge metode 3 , bilag II ) .

METHOD 2: DETERMINATION OF MOISTURE

METODE 2 : BESTEMMELSE AF VANDINDHOLD ( 102 * C OVN )

(oven 102 oC) .

1 . OMFANG OG ANVENDELSESOMRAADE

1. SCOPE AND FIELD OF APPLICATION

Denne metode anvendes ved bestemmelse af vandindhold :

This method determines the loss of mass on drying of:

- maelkepulver med hoejt fedtindhold ,

- dried high fat milk or high fat milk powder,

- maelkepulver eller soedmaelkspulver ,

- dried whole milk or whole milk powder,

- Pulver af delvis skummet maelk ,

- dried partly skimmed milk or partly skimmed-milk powder,

- skummetmaelkspulver .

- dried skimmed milk or skimmed-milk powder.

2 . DEFINITION

2. DEFINITION

Vandindhold = vaegttabet ved toerring bestemt som specificeret i metoden .

Moisture content: the loss of mass on drying as determined by the method specified.

3 . PRINCIP

3. PRINCIPLE

Restvaegten af en proevemaengde bestemmes efter toerring ved atmosfaerisk tryk i en ovn ved 102 mere aller mindre 1 * C til konstant vaegt . Vaegttabet beregnes som en vaegtprocent af proeven .

The residual mass of a test portion is determined after drying at atmospheric pressure in an oven at 102 oC ± 1 oC to constant mass. The loss of mass is calculated as a percentage by mass of the sample.

4 . APPARATUR

4. APPARATUS

4.1 . Analysevaegt .

4.1. Analytical balance.

4.2 . Skaale , fortrinsvis af nikkel , aluminium , rustfrit staal eller glas . Skaalene skal have laag , der passer godt , men som let kan fjernes . Passende dimensioner er : diameter 60 - 60 mm og dybde ca . 25 mm .

4.2. Dishes, preferably of nickel, aluminium, stainless steel or glass. The dishes must have lids which fit very well but which can readily be removed. Suitable dimensions are: diameter 60 to 80 mm and depth about 25 mm.

4.3 . Toerreovn ved atmosfaeretryk , godt ventileret , termostatreguleret med temperaturen reguleret til 102 mere aller mindre 1 * C . Temperaturen skal vaere den samme i hele ovnen .

4.3. Atmospheric-pressure drying oven, well ventilated, thermostatically controlled with temperature regulation (at 102 oC ± 1 oC). The temperature should be uniform throughout the oven.

4.4 . Ekssikkator , indeholdende friskt aktiveret silicagel med en indikator for vandindhold eller et tilsvarende toerremiddel .

4.4. Desiccator, containing freshly activated silica gel with a water content indicator or an equivalent desiccant.

5 . FREMGANGSMAADE

5. PROCEDURE

5.1 . Laaget tages af skaalen ( 4.2 . ) , og denne og laaget anbringes i ovnen ( 4.3 . ) og opvarmes i ca . en time .

5.1. Uncover the dish (4.2) and place it and its lid in the oven (4.3) and heat for about one hour.

5.2 . Laaget saettes paa skaalen , og den lukkede skaal flyttes over i ekssikkatoren ( 4.4 . ) . Man lader den afkoele til stuetemperatur , og der vejes med en noejagtighed paa 0,1 mg ( M0 ) .

5.2. Place the lid on the dish and transfer the covered dish to the desiccator (4.4). Allow it to cool to room temperature and accurately weigh to the nearest 0,1 mg (Mo).

5.3 . Ca . 2 g af pulverproeven overfoeres til skaalen , skaalen lukkes med laaget , og den lukkede skaal vejes saa hurtigt som muligt med en noejagtighed paa 0,1 mg ( M1 ) .

5.3. Introduce approximately 2 g of dried milk sample into the dish, cover the dish with the lid and accurately weigh to the nearest 0,1 mg the covered dish as quickly as possible (M1).

5.4 . Laaget tages af skaalen , og denne og laaget anbringes i ovnen i 2 timer .

5.4. Uncover the dish and put it with its lid in the oven for two hours.

5.5 . Laaget saettes paa igen , den lukkede skaal overflyttes til ekssikkatoren ; man lader den afkoele til stuetemperatur , og der vejes saa hurtigt som muligt med en noejagtighed paa 0,1 mg .

5.5. Replace the lid, transfer the covered dish to the desiccator, allow it to cool to room temperature and accurately weigh to the nearest 0,1 mg as quickly as possible.

5.6 . Laaget tages af skaalen , og denne og laaget opvarmes i 1 time i ovnen .

5.6. Uncover the dish and heat it and its lid for one hour in the oven.

5.7 . Procedure 5.5 . gentages .

5.7. Repeat process 5.5.

5.8 . Procedurerne 5.6 . og 5.5 . gentages indtil faldet i vaegten mellem to paa hinanden foelgende vejninger ikke overstiger 0,5 mg eller indtil vaegten stiger . Hvis der forekommer en stigning i vaegten , anvendes den laveste vaegt , der er opnaaet , ved beregningerne ( 6.1 . ) . Sidstnaevnte vaegt benaevnes M2 g .

5.8. Repeat processes 5.6 and 5.5 until the decrease in mass between the successive weighings does not exceed 0,5 mg or until the mass increases. If an increase in mass occurs use the lowest mass obtained in the calculation (6.1). Let the final weight recorded be M2 g.

6 . PRAESENTATION AF RESULTATERNE

6. EXPRESSION OF RESULTS

6.1 . Beregningsmetode

6.1. Method of calculation

Vaegttabet beregnes ved toerringen af proeven , udtrykt som en vaegtprocent ved hjaelp af foelgende formel :

Calculate the loss of mass on drying of the sample, expressed as a percentage by mass, by the formula:

( M1 - M2 ) / ( M1 - M0 ) gange 100

×100

hvor :

where:

M0 = vaegten i g af skaalen og dens laag efter procedure 5.2 . ;

M0 = mass, in g of the dish and its lid after process 5.2;

M1 = vaegten i g af skaalen , dens laag og proeven efter procedure 5.3 . ;

M1 = mass, in g of the dish, its lid and sample after process 5.3;

M2 = vaegten i g af skaalen , dens laag og slutproeven efter procedure 5.5 .

M2 = mass, in g of the dish, its lid and final sample after process 5.5.

6.2 . Repetitionsevne

6.2. Repeatability

Forskellen mellen resultaterne for to bestemmelser udfoert samtidigt eller hurtigt efter hinanden af den samme analytiker der anvender den samme proeve under de samme betingelser , maa ikke overstige 0,1 g vand pr . 100 g produkt .

The difference in results between two determinations carried out simultaneously or in rapid succession on the same sample, by the same analyst, under the same conditions, shall not exceed 0,1 g of moisture per 100 g of product.

METODE 3 : BESTEMMELSE AF FEDTINDHOLD ( ROESE-GOTTLIEB METODEN )

METHOD 3: DETERMINATION OF FAT CONTENT IN CONDENSED MILKS (RÖSE-GOTTLIEB METHOD)

1 . OMFANG OG ANVENDELSESOMRAADE

1. SCOPE AND FIELD OF APPLICATION

Denne metode anvendes ved bestemmelse af fedtindholdet i :

This method determines the fat content of:

- kondenseret maelk med hoejt fedtindhold eller usukret kondenseret maelk med hoejt fedtindhold ,

- unsweetened condensed high fat milk,

- kondenseret maelk , usukret kondenseret maelk eller kondenseret soedmaelk ,

- unsweetened condensed milk,

- kondenseret delvis skummet maelk eller usukret kondenseret delvis skummet maelk ,

- unsweetened condensed partly skimmed milk,

- kondenseret skummetmaelk eller usukret kondenseret skummetmaelk ,

- unsweetened condensed skimmed milk,

- sukret kondenseret maelk eller sukret kondenseret soedmaelk ,

- sweetened condensed milk,

- sukret kondenseret delvis skummet maelk ,

- sweetened condensed partly skimmed milk,

- sukret kondenseret skummetmaelk .

- sweetened condensed skimmed milk.

2 . DEFINITION

2. DEFINITION

Fedtindholdet i kondenseret maelk : fedtindholdet som det bestemmes ved den specificerede metode .

The fat content of condensed milks: fat content as determined by the method specified.

3 . DEFINITION

3. PRINCIPLE

Fedtindholdet bestemmes ved ekstraktion af fedtet fra en ammoniakalsk alkoholisk oploesning af proeven med diethylether og petroleumsether efterfulgt af afdampning af oploesningsmidlerne og vejning af resten og beregning som en vaegtprocent af proeven , svarende til Roese-Gottliebs princip .

The fat content is determined by extraction of the fat from an ammoniacal alcoholic solution of the sample with diethyl ether and light petroleum followed by evaporation of the solvents and weighing of the residue and calculation as a percentage by mass of the sample, according to the principle of Rose-Gottlieb.

4 . REAGENSER

4. REAGENTS

Alle reagenser skal opfylde de krav , der er specificeret i blindproeven ( 6.1 . ) Om noedvendigt maa oploesningsmidlerne redestilleres under tilstedevaerelsen af ca . 1 g smoerfedt pr . 100 ml oploesningsmiddel .

All reagents should conform to the requirements specified in the blank test (6.1). If necessary, reagents may be redistilled in the presence of about 1 g of butterfat for 100 ml of solvent.

4.1 . Ammoniakoploesning , ca . 25 % ( m/m ) NH3 ( vaegtfylde ved 20 * C ca . 0,91 g/ml ) , eller en staerkere oploesning af kendt koncentration .

4.1. Ammonia solution, approximately 25 % (m/m) NH3 (density at 20 oC approximately 0.91 g/ml), or a stronger solution of known concentration.

4.2 . Ethanol , 96 mere aller mindre 2 % ( v/v ) eller , hvis dette ikke er tilgaengeligt , ethanol denatureret med methanol , ethylmethylketon eller petroleumsether .

4.2. Ethanol, 96 ± 2 % (v/v) or, if not available, ethanol denatured with methanol, ethyl methyl ketone or light petroleum.

4.3 . Diethylether , peroxidfri .

4.3. Diethyl ether, peroxide-free.

Note 1 :

Note 1:

For at undersoege for peroxider , tilsaettes til 10 ml ether i et lille cylinderglas med glasprop og forskyllet med ether , 1 ml frisk fremstillet 10 % kaliumjodidoploesning . Blandingen rystes og henstilles i 1 minut . Der maa ikke kunne ses gul farve i noget af lagene .

To test for peroxides, add to 10 ml of the ether in a small glass stoppered cylinder, previously rinsed with the ether, 1 ml freshly prepared 10 % potassium iodide solution. Shake and let stand for one minute. No yellow colour should be observed in either layer.

Note 2 :

Note 2:

Diethylether kan holdes fri for peroxider ved at tilsaette vaadt zinkfolie , der har vaeret fuldstaendigt neddyppet i en fortyndet sur kobbersulfatoploesning i 1 min . og derefter vasket med vand . Der anvendes ca . 8 000 mm2 zinkfolie pr . liter ; det klippes i strimler , der er lange nok til at naa mindst halvt op i beholderen .

Diethyl ether may be maintained free from peroxides by adding wet zinc foil that has been completely immersed in dilute acidified copper sulphate solution for one minute and subsequently washed with water. Use per litre approximately 8000 mm2 zinc foil; cut in strips long enough to reach at least halfway up the container.

4.4 . Petroleumsether med kogepunkt mellem 30 og 60 * C .

4.4. Light petroleum (petroleum ether), with any boiling range between 30 and 60 oC.

4.5 . Blandet oploesningsmiddel , fremstillet kort foer brugen ved at blande lige store rumfang af diethylether ( 4.3 . ) og petroleumsether ( 4.4 . ) ( hvor der er anfoert brug af blandet oploesningsmiddel , kan det erstattes af enten diethylether eller petroleumsether alene ) .

4.5. Mixed solvent, prepared shortly before use by mixing equal volume of diethyl ether (4.3) and light petroleum (4.4) (where the use of mixed solvent is indicated, it may be replaced by either diethyl ether or light petroleum alone).

5 . APPARATUR

5. APPARATUS

5.1 . Analysevaegt

5.1. Analytical balance.

5.2 . Passende ekstraktionsroer eller kolber , udstyret med glasslibpropper , eller andet lukkeudstyr , der ikke paavirkes af de anvendte oploesningsmidler .

5.2. Suitable extraction tubes or flasks, provided with ground glass stoppers or other closures un-. affected by the solvents used.

5.3 . Kolber , tyndvaeggede og fladbundede , rumfang paa 150 - 250 ml .

5.3. Flasks, thin-walled and flat-bottomed, 150 to 250 ml capacity.

5.4 . Toerreovn ved atmosfaeretryk , godt ventileret og termostatreguleret ( indstillet til 102 mere aller mindre 1 * C ) .

5.4. Atmospheric pressure drying oven, well ventilated and thermostatically controlled (adjusted to operate at 102 oC ± 1 oC.

5.5 . Antistoedkogningskorn , fedtfrie , ikke poroese , ikke-smuldrende ved anvendelsen , f.eks . smaa glaskugler eller stykker af siliciumcarbid ( anvendelsen af dette materiale er valgfrit ; se paragraf 6.2.1 . ) .

5.5. Anti-bumping granules, fat-free, non porous, non friable in use, e.g. glass beads or pieces of silicon carbide (the use of this material is optional; see clause 6.2.1.

5.6 . Haevert , der passer til ekstraktionsroererne .

5.6. Siphon, to fit extraction tubes.

5.7 . Centrifuge

5.7. Centrifuge (optional).

6 . FREMGANGSMAADE

6. PROCEDURE

6.1 . Blind proeve

6.1. Blank test

Samtidig med bestemmelsen af fedtindholdet udfoeres en blindbestemmelse paa 10 ml vand , idet der anvendes samme type af ekstraktionsudstyr , de samme reagenser i de samme maengder og den samme fremgangsmaade , bortset fra paragraf 6.2.2 . Hvis blindproeven overstiger 0,5 mg , skal reagenserne undersoeges og de(t ) urene reagens(er ) skal renses eller udskiftes .

At the same time as the determination of the fat content of the sample, carry out a blank determination on 10 ml of water using the same type of extraction apparatus, the same reagents in the same amounts and the same procedure as described hereafter, excluding clause 6.2.2. If the blank exceeds 0.5 mg, the reagents should be checked and the impure reagent or reagents should be purified or replaced.

6.2 . Bestemmelse

6.2. Determination

6.2.1 . En kolbe ( 5.3 . ) ( samt , hvis det er noedvendigt , nogle antistoedkogningskorn ( 5.5 . ) til at sikre en langsom kogning ved den efterfoelgende fjernelse af oploesningsmidler ) toerres i ovnen ( 5.4 . ) i 1/2 - 1 time . Man lader kolben afkoele til temperaturen i vejerummet og den afkoelede kolbe vejes med en noejagtighed paa 0,1 mg .

6.2.1. Dry a flask (5.3) (together with, if required, some anti-bumping granules (5.5) to promote gentle boiling during the subsequent removal of the solvents) in the oven (5.4) for half to one hour. Allow the flask to cool to the temperature of the balance room and accurately weigh the cooled flask to the nearest 0,1 mg.

6.2.2 . Den fremstillede proeve paa 5 g omroeres og der afvejes straks 2 - 2,5 g proeve ( eller 4 - 5 g , naar det drejer sig om skummetmaelksprodukter ) med en noejagtighed paa 1 mg direkte , eller ved difference , i ektraktionsapparaturet ( 5.2 . ) . Der tilsaettes vand op til 10,5 ml og rystes forsigtigt under svag opvarmning ( 40 - 50 * C ) , indtil produktet er fuldstaendigt dispergeret . Proeven skal vaere fuldstaendigt dispergeret , i modsat fald maa bestemmelsen gentages .

6.2.2. Stir the prepared sample and immediately weigh, to the nearest 1 mg, 2 to 2,5 g of the sample if sweetened or 4 to 5 g of the sample if unsweetened directly in, or by difference into, the extraction apparatus (5.2). Add water to 10,5 ml and shake gently with slight warming (40 to 50 oC) until the product is completely dispersed. The sample must be dispersed completely otherwise the determination should be repeated.

6.2.3 . Der tilsaettes 1,5 ml ammoniak ( 25 % ) ( 4.1 . ) eller en tilsvarende maengde af en staerkere oploesning og der blandes omhyggeligt .

6.2.3. Add 1,5 ml ammonia (25 %) (4.1) or a corresponding volume of a stronger solution, and mix well.

6.2.4 . Der tilsaettes 10 ml ethanol ( 4.2 . ) og vaeskerne blandes forsigtigt , men omhyggeligt i det ulukkede apparatur .

6.2.4. Add 10 ml ethanol (4.2) and mix the liquids gently but thoroughly in the unclosed apparatus.

6.2.5 . Der tilsaettes 25 ml diethylether ( 4.3 . ) og der afkoeles under rindende vand . Apparaturet lukkes og rystes kraftigt , og det vendes gentagne gange i 1 minut .

6.2.5. Add 25 ml diethyl ether (4.3). Cool under running water. Close the apparatus and shake vigorously and invert repeatedly for one minute.

6.2.6 . Proppen fjernes forsigtigt , og der tilsaettes 25 ml petroleumsether ( 4.4 . ) , idet de foerste faa mililiter anvendes til at skylle proppen og den indvendige side af apparatets krave med , og skyllevaesken faar lov til at loebe ned i apparaturet . Der lukkes ved at saettes proppen paa igen , og apparatet rystes og vendes gentagne gange i 30 sek . Der maa ikke rystes for voldsomt , hvis der ikke anvendes centrifugering under 6.2.7 .

6.2.6. Remove the stopper carefully and add 25 ml light petroleum (4.4) using the first few millilitres to rinse the stopper and inside of the neck of the apparatus, allowing the rinsings to run into the apparatus. Close by replacing the stopper and shake and invert repeatedly for 30 seconds. Do not shake too vigorously if centrifuging is not to be used in 6.2.7.

6.2.7 . Apparaturet henstilles , indtil det oeverste lag er blevet klart og det tydeligt har adskilt sig fra det underste lag . Alternativt kan separeringen ske ved hjaelp af en centrifuge af passende art ( 5.7 . )

6.2.7. Allow the apparatus to stand until the upper liquid layer has become clear and has distinctly separated from the lower aqueous layer. Alternatively carry out the separation using a suitable centrifuge (5.7).

Note :

Note:

Hvis der anvendes en centrifuge , der ikke drives af en trefaset motor , kan der forekomme gnistdannelse , og der maa udvises forsigtighed for at undgaa en eksplosion eller brand af etherdamp , der f.eks . kommer fra et ituslaaet centrifugeglas .

When a centrifuge which is not driven by a three-phase motor, is used, sparks may occur and care must therefore be taken to avoid an explosion or fire from any ether vapours, coming, for example, from a broken tube.

6.2.8 . Proppen fjernes , og denne og indersiden af apparaturets krave skylles med faa ml blandet oploesningsmiddel ( 4.5 . ) ; skyllevaesken faar lov til at loebe ned i apparaturet . Saa meget som muligt at det oeverste lag overfoeres omhyggeligt enten ved dekantering eller ved hjaelp af en haevert ( 5.6 . ) til den forberedte kolbe ( 6.2.1 . ) .

6.2.8. Remove the stopper, rinse it and the inside of the neck of the apparatus with a few millilitres of mixed solvent (4.5) and allow the rinsings to run into the apparatus. Carefully transfer as much as possible of the supernatant layer by decantation or by means of a siphon (5.6) into the pre pared flask (6.2.1).

Note :

Note:

Hvis overfoerselen ikke sker ved hjaelp af en haevert , kan det vaere noedvendigt at tilsaette en smule vand for at haeve skillefladen mellem de to lag og saaledes lette dekanteringen .

If the transfer is not made using a siphon, it may be necessary to add a little water in order to raise the interface between the two layers thus aiding decantation.

6.2.9 . Ydersiden og indersiden af kraven paa apparaturet eller spidsen af haeverten skylles med faa ml blandet oploesningsmiddel . Skyllevaeskerne fra ydersiden af apparaturet faar lov til at loebe ned i kolben , og skyllevaesken fra undersiden af kraven og fra haevertens spids loeber ned i ekstraktionsapparaturet .

6.2.9. Rinse the outside and the inside of the neck of the apparatus or the tip and the lower part of the siphon with a few millilitres of mixed solvent (4.5). Allow the rinsings from the outside of the apparatus to run into the flask and the rinsings from the inside of the neck and from the siphon to run into the extraction apparatus.

6.2.10 . Ekstraktion nummer to foretages ved at gentage procedurerne fra 6.2.5 . til 6.2.9 . inklusive , men der anvendes kun 15 ml diethylether og 15 ml petroleumsether .

6.2.10. Make a second extraction by repeating the procedure of 6.2.5 to 6.2.9 inclusive but using only 15 ml diethyl ether and 15 ml light petroleum.

6.2.11 . En tredje ekstraktion foretages ved at gentage procedure 6.2.10 . , men den sidste skylning ( 6.2.9 . ) , udelades .

6.2.11. Make a third extraction by repeating the procedure of 6.2.10 but omit the final rinsing (6.2.9).

Note :

Note:

Det er ikke paakraevet at udfoere denne tredje ekstraktion , naar der analyseres paa usukret kondenseret skummetmaelk og sukret kondenseret skummet maelk .

It is not mandatory to carry out this third extraction when analysing skimmed unsweetened condensed milk and skimmed sweetened condensed milk samples.

6.2.12 . Saa meget oploesningsmiddel ( inklusive ethanol ) som muligt afdampes eller afdestilleres forsigtigt . Hvis kolbens rumfang er for lille , vil det vaere noedvendigt at fjerne noget af oploesningsmidlet som ovenfor efter hver ekstraktion .

6.2.12. Carefully evaporate or distil off as much solvent (including the ethanol) as possible. If the flask is of small capacity, it will be necessary to remove some of the solvent as above after each extraction.

6.2.13 . Naar der ikke laengere er tydelig lugt af oploesningsmiddel , placeres kolben paa siden i ovnen og opvarmes i en time .

6.2.13. When there is no appreciable odour of solvent place the flask on its side in the oven and heat for one hour.

6.2.14 . Kolben fjernes fra ovnen , man lader den afkoele til vejerummets temperatur , og den vejes med en noejagtighed paa 0,1 mg .

6.2.14. Remove the flask from the oven, allow to cool to the temperature of the balance room and accurately weigh to the nearest 0,1 mg.

6.2.15 . Gentag 6.2.15 . og 6.2.14 . med varmeperioder paa 30 - 60 min . , indtil faldet i vaegten mellem to paa hinanden foelgende vejninger ikke overstiger 0,5 mg eller indtil vaegten stiger . Hvis der forekommer en stigning i vaegten , anvendes den laveste vaegt , der er opnaaet , ved beregningerne ( 7.1 . ) . Sidstnaevnte vaegt benaevnes M1 g .

6.2.15. Repeat 6.2.13 and 6.2.14 for heating periods of 30 to 60 minutes until the difference in mass of two successive weighings is less than 0.5 mg or until the mass increases. If an increase in mass occurs use the lowest mass obtained in the calculation (7.1). Let the final weight recorded be M1 g.

6.2.16 . 15 - 25 ml petroleumsether tilsaettes for at bekraefte , at det ekstraherede materiale er helt oploeseligt . Der opvarmes forsigtigt , og oploesningsmidlet drejes rundt , indtil alt fedt er oploest .

6.2.16. Add 15 to 25 ml light petroleum in order to confirm that the extracted matter is wholly soluble. Warm gently and swirl the solvent until all the fat is dissolved.

6.2.16.1 . Hvis det ekstraherede materiale er helt oploeseligt i petroleumsether , er vaegten af fedtet differencen mellem vaegtene , der er bestemt i 6.2.1 . og 6.2.15 .

6.2.16.1. If the extracted matter is wholly soluble in the light petroleum, the mass of fat is the difference between the weights determined at stages 6.2.1 and 6.2.15.

6.2.16.2 . Hvis der er uoploeseligt materiale til stede , eller saafremt der er tvivl ekstraheres fedtet fuldstaendigt fra kolben ved gentagne udvaskninger med varm petroleumsether , idet man lader det uoploeste materiale bundfaelde sig inden hver dekantering . Ydersiden af kolbens krave skylles tre gange . Kolben opvarmes placeret paa siden i 1 time i ovnen , man lader den afkoele til vejerummets temperatur som foer ( 6.2.1 ) , og den vejes med en noejagtighed paa 0,1 mg . Vaegten af fedtet er differencen mellem vaegten opnaaet i 6.2.15 . og denne sidste vaegt .

6.2.16.2. If any insoluble matter is present, or in case of doubt, completely extract the fat from the flasks by repeated washing with warm light petroleum, allowing the undissolved material to settle before each decantation. Rinse the outside of the neck of the flask three times. Heat the flask, placed on its side, for one hour in the oven, allow to cool to the temperature of the balance room as before (6.2.1) and weigh to the nearest 0,1 mg. The mass of fat is the difference between the mass obtained at 6.2.15 and this final mass.

7 . PRAESENTATION AF RESULTATERNE

7. EXPRESSION OF RESULTS

7.1 . Beregning

7.1. Calculation

Vaegten i g af ekstraheret fedt er :

The mass, in g of fat extracted is:

( M1 - M2 ) - ( B1 - B2 )

(M1 — M2) — (B1 — B2)

og fedtindholdet i proeven , udtrykt som procentdel er :

and the fat content of the sample, expressed as a percentage is:

( ( M1 - M2 ) - ( B1 - B2 ) ) /S gange 100

×100

hvor :

where:

M1 = vaegten i g af kolben M med fedt efter trin 6.2.15 ;

M1 = mass, in g of flask M with fat after stage 6.2.15;

M2 = vaegten i g af kolben M efter trin 6.2.1 . eller i tilfaelde af uoploest materiale eller tvivl trin 6.2.16.2 ;

M2 = mass, in g of flask M after stage 6.2.1 or, in the case of undissolved material or doubt, stage 6.2.16.2;

B1 = vaegten i g af kolben B ved blindvaerdien efter trin 6.2.15 ;

B1 = mass, in g of flask B of the blank after stage 6.2.15;

B2 = vaegten i g af kolben B efter trin 6.2.1 . eller i tilfaelde af uoploest materiale eller tvivl efter trin 6.2.16.2 ;

B2 = mass, in g of flask B after stage 6.2.1 or, in the case of undissolved material or doubt, stage 6.2.16.2;

S = vaegten i g af anvendt proeve .

S = mass, in g of sample used.

7.2 . Repetitionsevne

7.2. Repeatability

Forskellen mellem resultaterne for to bestemmelser udfoert samtidigt eller hurtigt efter hinanden af den samme analytiker , der anvender den samme proeve , maa ikke overstige 0,05 g fedt pr . 100 g produkt .

The difference between results of two determinations carried out obtained simultaneously or in rapid succession on the same sample, by the same analyst, under the same conditions, shall not exceed 0,05 g fat per 100 g of the product.

METODE 4 : BESTEMMELSE AF FEDTINDHOLD ( ROESE-GOTTLIEB METODE )

METHOD 4: DETERMINATION OF FAT CONTENT IN DRIED MILKS (RÖSE-GOTTLIEB METHOD)

1 . OMFANG OG ANVENDELSESOMRAADE

1. SCOPE AND FIELD AND APPLICATION

Denne metode anvendes ved bestemmelse af fedtindholdet i :

This method determines the fat content of:

- maelkepulver med hoejt fedtindhold ,

- dried high fat milk or high fat milk powder,

- maelkepulver eller soedmaelkspulver ,

- dried whole milk or whole milk powder,

- pulver af delvis skummet maelk ,

- dried partly skimmed milk or partly skimmed-milk powder,

- skummetmaelkspulver .

- dried skimmed milk or skimmed-milk powder.

2 . DEFINITION

2. DEFINITION

Fedtindholdet i maelkepulver : fedtindholdet som det bestemmes ved den specificerede metode .

The fat content of dried milks: fat content as determined by the method specified.

3 . PRINCIP

3. PRINCIPLE

The fat content is determined by extraction of the fat from an ammoniacal alcoholic solution of sample with diethyl ether and light petroleum, followed by evaporation of the solvents and weighing of the residue and calculation as a percentage by mass of the sample, according to the principle of Rose-Gottlieb.

4 . REAGENSER

4. REAGENTS

Alle reagenser skal opfylde de krav , der er specificeret i blindproeven ( 6.1 . ) . Om noedvendigt maa reagenserne redestilles under tilstedevaerelsen af ca . 1 g smoerfedt pr . 100 ml oploesningsmiddel .

All reagents should conform to the requirements specified in the blank test (6.1). If necessary, reagents may be redistilled in the presence of about 1 g of butterfat per 100 ml of solvent.

4.1 . Ammoniakoploesning , ca . 25 % ( m/m ) NH3 ( vaegtfylde ved 20 * C ca . 0,91 g/ml ) , eller en staerkere oploesning af kendt koncentration .

4.1. Ammonia solution, approximately 25 % (m/m) NH3 (density at 20 oC approximately 0.91 g/ml), or stronger solution of known concentration.

4.2 . Ethanol , 96 mere aller mindre 2 % ( v/v ) eller , hvis dette ikke er tilgaengeligt , ethanol denatureret med methanol , ethylmethylketon eller petroleumsether .

4.2. Ethanol, 96 ± 2 % (v/v) or, if not available, ethanol denatured with methanol, ethyl methyl ketone or light petroleum.

4.3 . Diethylether , peroxidfri .

4.3. Diethyl ether, peroxide-free

Note 1 :

Note 1:

For at undersoege for peroxider tilsaettes til 10 ml ether i et lille cylinderglas lukket med glasprop og forskyllet med ether , 1 ml frisk fremstillet 10 % kaliumjodidoploesning . Blandingen rystes og henstilles i 1 minut . Der maa ikke kunne ses gul farve i noget af lagene .

To test for peroxide, add to 10 ml of the ether in a small glass stoppered cylinder, previously rinsed with the ether, 1 ml freshly prepared 10 % potassium iodide solution. Shake and let stand for one minute. No yellow colour should be observed in either layer.

Note 2 :

Note 2:

Diethylether kan holdes fri for peroxider ved at tilsaette vaadt zinkfolie , der har vaeret fuldstaendigt neddyppet i en fortyndet sur kobbersulfatoploesning i 1 min . og derefter med vand . Der anvendes ca . 8 000 mm2 zinkfolie pr . liter ; det klippes i strimler , der er lange nok til at naa mindst halvt op i beholderen .

Diethyl ether may be maintained free from peroxides by adding wet zinc foil that has been completely immersed in dilute acidified copper sulphate solution for one minute and subsequently washed with water. Use per litre approximately 8000 mm2 zinc foil cut in strips long enough to reach at least halfway up the container.

4.4 . Petroleumsether med kogepunkt mellem 30 og 60 * C .

4.4. Light petroleum (petroleum ether), with any boiling range between 30 and 60 oC.

4.5. Mixed solvent, prepared shortly before use by mixing equal volumes of diethyl ether (4.3) and light petroleum (4.4) (when the use of mixed solvent is indicated, it may be replaced by either diethyl ether or light petroleum alone).

5 . APPARATUR

5. APPARATUS

5.1 . Analysevaegt

5.1. Analytical balance.

5.2 . Passende ekstraktionsroer eller kolber , udstyret med glasslibpropper , eller andet lukkeudstyr , der ikke paavirkes af de anvendte oploesningsmidler .

5.2. Suitable extraction tubes or flasks, provided with ground glass stoppers or other closures unaffected by the solvents used.

5.3 . Kolber , tyndvaeggede og fladbundede , rumfang paa 150 - 250 ml .

5.3. Flasks, thin-walled, flat-bottomed, of 150 to 250 ml capacity.

5.4 . Toerreovn ved atmosfaeretryk , godt ventileret og termostatreguleret ( indstillet til 102 mere aller mindre 1 * C ) .

5.4. Atmospheric pressure drying oven, well ventilated and thermostatically controlled (adjusted to operate at 102 oC ± 1 oC).

5.5. Anti-bumping granules, fat-free, non porous, non friable in use, e.g. glass beads or pieces of silicon carbide (the use of this material is optional: see clause 6.2.1).

5.6 . Vandbad , ved 60 - 70 * C .

5.6. Waterbath, at 60 to 70 oC.

5.7 . Haevert , der passer til ekstraktionsroerene .

5.7. Siphon to fit extraction tubes.

5.8 . Centrifuge

5.8. Centrifuge (optional).

6 . FREMGANGSMAADE

6. PROCEDURE

6.1 . Blindproeve

6.1. Blank test

Samtidig med bestemmelsen af fedtindholdet i proeven udfoeres en blindbestemmelse paa 10 ml vand , idet der anvendes samme type af ekstraktionsudstyr , de samme reagenser i de samme maengder og den samme fremgangsmaade , bortset fra paragraf 6.2.2 . Hvis blindproeven overstiger 0,5 mg , skal reagenserne undersoeges og de(t ) urene reagens(er ) skal renses eller udskiftes .

At the same time as the determination of the fat content of the sample, carry out a blank determination on 10 ml of water using the same type of extraction apparatus, the same reagents in the same amounts and the same procedure as described hereafter, excluding clause 6.2.2. If blank exceeds 0.5 mg, the reagents should be checked and the impure reagent or reagents should be purified or replaced.

6.2 . Bestemmelse

6.2. Determination

6.2.1 . En kolbe ( 5.3 . ) ( samt , hvis det er noedvendigt nogle antistoedkogningskorn ( 5.5 . ) til at sikre en langsom kogning ved den efterfoelgende fjernelse af oploesningsmidler ) toerres i ovnen ( 5.4 . ) i 1/2 - 1 time . Man lader kolben afkoele til temperaturen i vejerummet og den afkoelede kolbe vejes med en noejagtighed paa 0,1 mg .

6.2.1. Dry the flask (5.3) together with, if required, some anti-bumping granules (5.5) to promote gentle boiling during the subsequent removal of the solvents) in the oven (5.4) for half to one hour. Allow the flask to cool to the temperature of the balance room and accurately, weigh the cooled flask to the nearest 0,1 mg.

6.2.2 . Ca . 1 g soedmaelkspulver eller ca . 1,5 g pulver af delvis skummet maelk eller skummetmaelkspulver afvejes direkte i eller ved difference i ekstraktionsapparatet ( 5.2 . ) med en noejagtighed paa 1 mg . 10 ml vand tilsaettes , og der rystes forsigtigt , indtil maelkepulveret er fuldstaendigt dispergeret ( det kan vaere noedvendigt at opvarme nogle proever ) .

6.2.2 Accurately weigh, to the nearest 1 mg, directly in, or by difference into, the extraction apparatus (5.2) about 1 g of whole milk powder or about 1,5 g of partly skimmed or skimmed-milk powder. Add 10 ml water and shake gently until the milk powder is completely dispersed (heat may be necessary for some samples).

6.2.3 . 1,5 ml ammoniak ( 25 % ) ( 4.1 . ) eller en tilsvarende maengde af en staerkere oploesning tilsaettes , og der opvarmes paa et vandbad i 15 min . ved 60 - 70 * C , idet der lejlighedsvis omrystes . Der afkoeles , f.eks . under rindende vand .

6.2.3. Add 1.5 ml ammonia (25 %) (4.1) or a corresponding volume of a stronger solution and heat in a waterbath (5.6) for 15 minutes at 60 to 70 oC, shaking occasionally. Cool, for example, in running water.

6.2.4 . 10 ml ethanol ( 4.2 . ) tilsaettes , og vaeskerne blandes forsigtigt men omhyggeligt i det ulukkede apparatur .

6.2.4. Add 10 ml ethanol (4.2) and mix the liquids gently but thoroughly in the unclosed apparatus.

6.2.5 . 25 ml diethylether ( 4.3 . ) tilsaettes . Der afkoeles under rindende vand . Apparaturet lukkes og rystes kraftigt ; det vendes gentagne gange i 1 minut .

6.2.5. Add 25 ml diethyl ether (4.3). Cool in running water. Close the apparatus and shake vigorously and invert repeatedly for one minute.

6.2.6 . Proppen fjernes forsigtigt , og der tilsaettes 25 ml petroleumsether ( 4.4 . ) , idet de foerste faa milliliter anvendes til at skylle proppen og den indvendige side af apparatets krave med , og skyllevaesken faar lov til at loebe ned i apparaturet . Der lukkes ved at saette proppen paa igen , og apparatet rystes og vendes gentagne gange i 30 sek . Der maa ikke rystes for voldsomt , hvis der ikke anvendes centrifugering under 6.2.7 .

Note :

Note:

Hvis der anvendes en centrifuge , der ikke drives af en trefaset motor , kan der forekomme gnistdannelse , og der maa udvises forsigtighed for at undgaa en eksplosion eller brand af etherdampe , der f.eks . kommer fra et ituslaaet centrifugeglas .

When a centrifuge which is not driven by a three-phase motor is used, sparks may occur and care must therefore be taken to avoid an explosion or fire from any ether vapours coming, for example, from a broken tube.

6.2.8 . Proppen fjernes , og denne og indersiden af apparaturets krave skylles med faa ml blandet oploesningsmiddel ( 4.5 . ) ; skyllevaesken faar lov til at loebe ned i apparaturet . Saa meget som muligt af det oeverste lag overfoeres omhyggeligt enten ved dekantering eller ved hjaelp af en haevert ( 5.7 . ) til den forberedte kolbe ( 6.2.1 . ) .

Note :

Note:

Hvis overfoerselen ikke sker ved hjaelp af en haevert , kan det vaere noedvendigt at tilsaette en smule vand for at haeve skillefladen mellem de to lag og saaledes lette dekanteringen .

If the transfer is not made using a siphon, it may be necessary to add a little water in order to raise the interface between the two layers thus aiding decantation.

6.2.9 . Ydersiden og indersiden af kraven paa apparaturet eller spidsen af haeverten skylles med faa ml blandet oploesningsmiddel . Skyllevaeskerne fra ydersiden af apparaturet faar lov til at loebe ned i kolben , og skyllevaesken fra indersiden af kraven og fra haevertens spids loeber ned i ekstraktionsapparaturet .

6.2.9. Rinse the outside and the inside of the neck of the apparatus or the tip and the lower part of the siphon with a few millilitres of mixed solvent. Allow the rinsings from the outside of the appara tus to run into the flask and the rinsings from the inside of the neck and from the siphon to run into the extraction apparatus.

6.2.10 . Ekstraktion nummer to foretages ved at gentage procedurerne fra 6.2.5 . til 6.2.9 . inklusive , men der anvendes kun 15 ml diethylether og 15 ml petroleumsether .

6.2.10. Make a second extraction by repeating the procedure of 6.2.5 to 6.2.9 inclusive but using only 15 ml diethyl ether and 15 ml light petroleum.

6.2.11 . En tredje ekstraktion foretages ved at gentage procedure 6.2.10 . , men den sidste skylning ( 6.2.9 . ) udelades .

6.2.11. Make a third extraction by repeating the procedure of 6.2.10 but omit the final rinsing (6.2.9).

Note :

Note:

Det er ikke paakraevet at udfoere denne tredje ekstraktion , naar analysen foretages paa skummetmaelkspulver .

It is not mandatory to carry out this third extraction when analysing dried skimmed milk samples.

6.2.12. Carefully evaporate or distil off as much solvent (including the ethanol) as possible. If the flask is of small capacity it will be necessary to remove some of the solvent as above after each extraction.

6.2.13 . Naar der ikke laengere er tydelig lugt af oploesningsmiddel , placeres kolben paa siden i ovnen , og den opvarmes i en time .

6.2.13. When there is no appreciable odour of solvent, place the flask on its side in the oven and heat for one hour.

6.2.14 . Kolben fjernes fra ovnen , man lader den afkoele til vejerummets temperatur som tidligere ( 6.2.1 . ) , og den vejes med en noejagtighed paa 0,1 mg .

6.2.14. Remove the flask from the oven, allow to cool to the temperature of the balance room as previously (6.2.1) and accurately weigh to the nearest 0,1 mg.

6.2.15 . Gentag operationerne 6.2.13 . og 6.2.14 . med varmeperioder paa 30 - 60 min . , indtil faldet i vaegten mellem to paa hinanden foelgende vejninger ikke overstiger 0,5 mg eller indtil vaegten stiger . Hvis der forekommer en stigning i vaegten , anvendes den laveste vaegt , der er opnaaet , ved beregningerne ( 7.1 . ) . Sidstnaevnte benaevnes M1 g .

6.2.15. Repeat 6.2.13 and 6.2.14 for heating periods of 30 to 60 minutes until the difference in mass of two successive weights is less than 0,5 mg or until the mass increases. If an increase in mass occurs use the lowest mass obtained in the calculation (7.1). Let the final weight recorded be M1 g.

6.2.16. Add 15 to 25 ml light petroleum in order to confirm that the extracted matter is wholly soluble. Warm gently and swirl the solvent until all the fat is dissolved.

6.2.16.1 . Hvis det ekstraherede materiale er helt oploeseligt i petroleumsether er vaegten af fedtet differencen mellem vaegtene , der er bestemt i 6.2.1 . og 6.2.15 .

6.2.16.1. If the extracted matter is wholly soluble in the light petroleum, the mass of fat is the difference between the weights determined at stages 6.2.1 and 6.2.15.

6.2.16.2. If any insoluble matter is present, or in case of doubt completely extract the fat from the flask by repeated washing with warm light petroleum, allowing the undissolved material to settle before each decantation. Rinse the outside of the neck of the flask three times.

Kolben opvarmes placeret paa siden i 1 time i ovnen , man lader den afkoele til vejerummets temperatur som foer ( 6.2.1 . ) , og den vejes med en noejagtighed paa 0,1 mg . Vaegten af fedtet er differencen mellem vaegten opnaaet i 6.2.15 . og denne sidste vaegt .

Heat the flask, placed on its side, for one hour in the oven, allow to cool to the temperature of the balance room, as before (6.2.1) and weigh to the nearest 0,1 mg. The mass of fat is the difference between the mass under 6.2.15 and this final mass.

7 . PRAESENTATION AF RESULTATERNE

7. EXPRESSION OF RESULTS

7.1 . Beregning

7.1. Calculation

Vaegten i g af ekstraheret fedt er :

The mass, in g of fat extracted is:

( M1 - M2 ) - ( B1 - B2 )

(M1 — M2) — (B1 — B2)

og fedtindholdet i proeven , udtrykt som procentdel er :

and the fat content of the sample, expressed as a percentage, is:

( M1 - M2 ) - ( B1 - B2 )/S gange 100

×100

hvor :

where:

M1 = vaegten i g af kolben M med fedt efter trin 6.2.15 ;

M1 = mass, in g of flask M with fat after stage 6.2.15;

M2 = vaegten i g af kolben M efter trin 6.2.1 . eller i tilfaelde af uoploest materiale eller tvivl trin 6.2.16.2 ;

M2 = mass, in g of flask M after stage 6.2.1 or, in the case of undissolved material or doubt, stage 6.2.16.2;

B1 = vaegten i g af kolben B ved blindvaerdien efter trin 6.2.15 ;

B1 = mass, in g of flask B of the blank after stage 6.2.15;

B2 = vaegten i g af kolben B efter trin 6.2.1 . eller i tilfaelde af uoploest materiale eller tvivl efter trin 6.2.16.2 ;

B2 = mass, in g of flask B after stage 6.2.1 or, in the case of undissolved material or doubt, stage 6.2.16.2;

S = vaegten i g af anvendt proeve .

S = mass, in g of sample used.

7.2 . Repetitionsevne

7.2. Repeatability

Forskellen mellem resultaterne for to bestemmelser udfoert samtidigt eller hurtigt efter hinanden af den samme analytiker der anvender den samme proeve under de samme betingelser , maa ikke overstige 0,2 g fedt pr . 100 g produkt , for skummetmaelkspulver dog 0,1 g fedt pr . 100 g produkt .

The difference between results of two determinations carried out simultaneously or in rapid succession on the same sample, the same analyst, under the same conditions, shall not exceed 0,2 g fat per 100 g of product with the exception of skimmed-milk powder for which the difference must not exceed 0,1 g fat per 100 g of product.

METODE 5 : BESTEMMELSE AF SACCHAROSEINDHOLD ( POLARIMETRISK METODE )

METHOD 5: DETERMINATION OF SUCROSE CONTENT (POLARIMETERIC METHOD)

1 . OMFANG OG ANVENDELSESOMRAADE

1. SCOPE AND FIELD OF APPLICATION

Denne metode anvendes ved bestemmelse af saccharoseindholdet i :

This method determines the sucrose content of:

- sukret kondenseret maelk eller sukret kondenseret soedmaelk ,

- sweetened condensed milk,

- sukret kondenseret delvis skummet maelk ,

- sweetened condensed partly skimmed milk,

- sukret kondenseret skummetmaelk .

- sweetened condensed skimmed milk.

Proeverne maa ikke indeholde invertsukker .

Samples must not contain invert sugar.

2 . DEFINITION

2. DEFINITION

Saccharoseindholdet i sukret kondenseret maelk : saccharoseindholdet som det bestemmes ved den specificerede metode .

The sucrose content of sweetened condensed milks: the sucrose content as determined by the method specified.

3 . PRINCIP

3. PRINCIPLE

Metoden er baseret paa Clerget inversionsprincippet , en mild behandling af proeven med syre , der giver en fuldstaendig hydrolyse af saccharose , men naesten ingen af lactose eller andre sukkerarter . Saccharoseindholdet opnaas ud fra aendringen i oploesningens drejningsevne .

The method is based on the principle of the Clerget inversion, a mild treatment of the sample with acid which produces complete hydrolysis of sucrose but almost none of lactose or other sugars. The sucrose content is obtained from the change in rotating power of the solution.

Et klart filtrat af oploesningen , uden mutarotation af lactose , fremstilles ved behandling med ammoniak efterfulgt af neutralisation og klaring ved efterfoelgende tilsaetning af zinkacetat - og kaliumhexacyanoferrat(II)-oploesninger .

A clear filtrate of the sample, without mutarotation by lactose, is prepared by treatment of the solution with ammonia followed by neutralization and clearing by the successive addition of zinc acetate and potassium hexacyanoferrate II solutions.

Paa en del af filtratet hydrolyseres saccharosen paa en specificeret maade .

In a portion of the filtrate the sucrose is hydrolyzed in a specified manner.

Ud fra rotationen af filtratet foer og efter inversionen kan saccharoseindholdet beregnes ved at anvende passende formler .

From the rotation of the filtrate before and after inversion, the sucrose content is calculated using the appropriate formulae.

4 . REAGENSER

4. REAGENTS

4.1 . Zinkacetatoploesning , M1 : 21,9 g krystallinsk zinkacetatdihydrat Zn(C2H3O2)3 * 2H2O og 3 ml iseddike oploeses i vand , og der fyldes op til 100 ml med vand .

4.1. Zinc acetate solution, 1 M: dissolve 21,9 g crystallized zinc acetate dihydrate Zn(C2H.O2)2.2H2O and 3 ml glacial acetic acid in water and make up to 100 ml with water.

4.2 . Kaliumhexacyanoferrat(II)oploesning , 0,25 M1 : 10,6 g krystallinsk kaliumhexacyanoferrat(II)-trihydrat K4(Fe(CN)6 ) * 3H2O oploeses i vand , og der fyldes op til 100 ml med vand .

4.2. Potassium hexacyanoferrate (II) solution, 0,25 M: dissolve 10,6 g crystallized potassium hexacyanoferrate (II) trihydrate K4[Fe(CN)6]. 3H2O in water and make up to 100 ml with water.

4.3 . Saltsyreoploesning , 6,35 mere aller mindre 0,20 M1 ( 20 - 22 % ) eller 5,0 mere aller mindre 0,2 mol * 1 -1 16 - 18 % ) .

4.3. Hydrochloric acid solution, 6,35 ± 0,20 M (20 to 22 %) or 5,0 ± 0,2 M (16 to 18 %).

4.4 . Ammoniakoploesning , 2,0 mere aller mindre 0,2 M1 ( 3,5 % ) .

4.4. Ammonia solution, 2,0 ± 0,2 M (3,5 %).

4.5 . Eddikesyreoploesning , 2,0 mere aller mindre 0,2 M1 ( 12 % ) .

4.5. Acetic acid solution, 2,0 ± 0,2 M (12 %).

4.6 . Bromothylmolblaat , indikator , 1 % ( m/v ) oploesning i ethanol .

4.6. Bromothymol blue indicator, 1 % (m/v) solution in ethanol.

5 . APPARATUR

5. APPARATUS

5.1 . Vaegt , med en foelsomhed paa 10 mg .

5.1. Balance, sensitivity 10 mg.

5.2 . Polarimeterroer , 2 dm , med noejagtig kalibreret laengde .

5.2. Polarimeter tube, 2dm, of exactly calibrated length.

5.3 . Polarimeter eller saccharimeter .

5.3. Polarimeter or saccarimeter:

a ) Polarimeter med natriumlys eller groent kviksoelvlys ( kviksoelvdamplampe med prisme eller den specielle Wratten Screen nr . 77 A ) , der kan aflaeses med en noejagtighed paa mindst 0,05 vinkelgrader .

(a) Polarimeter with sodium light or mercury green light (mercury vapour lamp with prism or the special Wratten Screen No 77 A), to be read with an accuracy of at least 0.05 angular degrees,

b ) Saccharimeter med international sukkerskala , der anvender hvidt lys , som passerer et filter paa 15 mm af en 6 % oploesning af kaliumdichromat , eller natriumlys , og som kan aflaeses med en noejagtighed paa mindst 0,1 * paa den internationale sukkerskala .

(b) Saccarimeter with international sugar scale, using white light passing through a filter of 15 mm of a 6 % solution of potassium bichromate, or sodium light, to be read with an accuracy of at least 0,1o on the international sugar scale.

5.4 . Vandbad , reguleret til 60 mere aller mindre 1 * C .

5.4. Water bath, regulated at 60 oC ± 1 oC.

6 . FREMGANGSMAADE

6. PROCEDURE

6.1 . Kontrolbestemmelse

6.1. Control determination

For at standardisere fremgangsmaaden , reagenserne og apparaturet udfoeres en dobbelt kontrolbestemmelse som anfoert nedenfor , idet der anvendes 100 g maelk og 18,00 g ren saccharose eller en blanding af 110 g skummetmaelk og 18,00 g ren saccharose , svarende til 40,00 g kondenseret maelk indeholdende 45 % saccharose . Sukkerindholdet beregnes ved at anvende formlerne i 7 , idet M , F og P i formel 1 erstattes med henholdsvis maengden af maelk , der er taget i brug , og fedt - og proteinindholdet i denne maelk , og i formel 2 indsaettes for M vaerdien 40,00 . Gennemsnittet af de fundne vaerdier maa ikke afvige mere end 0,2 % fra 45,0 % .

In order to standardize the procedure, reagents and apparatus, carry out a control determination in duplicate as described below using a mixture of 100 g of milk and 18 g pure sucrose or a mixture of 110 g of skimmed milk and 18 g pure sucrose, each corresponding to 40 g of condensed milk containing 45 % sucrose. Calculate the sugar content using the formulae under 7, substituting for M, F and P respectively in formula 1 the quantity of milk taken and the fat and protein content of this milk, and in formula 2 for M, the value of 40,00. The mean of the values found shall not differ by more than 0,2 % from 45,0 %.

6.2 . Bestemmelse

6.2. Determination

6.2.1 . Med en noejagtighed paa 10 mg afvejes 40 g af den godt blandede proeve i baegerglas paa 100 ml . 50 ml varmt vand ( 80 - 90 * C ) tilsaettes og der blandes godt .

6.2.1. Weigh to within 10 mg, approximately 40 g of the well mixed sample into a 100 ml glass beaker. Add 50 ml of hot water (80 to 90 oC) and mix well.

6.2.2 . Blandingen overfoeres kvantitativt til en 200 ml maalekolbe , idet baegerglasset skylles gentagne gange med 60 * C varmt vand , indtil rumfanget er 120 - 150 ml . Der blandes og afkoeles til stuetemperatur .

6.2.2. Transfer the mixture quantitatively to a 200 ml measuring flask, rinsing the beaker with successive quantities of water at 60 oC, until the total volume is between 120 and 150 ml. Mix and cool to room temperature.

6.2.3 . Der tilsaettes 5 ml fortyndet ammoniakoploesning ( 4.4 . ) . Der blandes igen , og blandingen henstilles derefter i 15 min .

6.2.3. Add 5 ml of the dilute ammonia solution (4.4). Mix again and then allow to stand for 15 minutes.

6.2.4 . Ammoniakken neutraliseres ved at tilsaette en aekvivalent maengde af den fortyndede eddikesyreoploesning ( 4.5 . ) . Det noejagtige antal ml bestemmes paa forhaand ved at titrere ammoniakoploesningen , idet bromothylmolblaat ( 4.6 . ) anvendes som indikator . Der blandes .

6.2.4. Neutralize the ammonia by adding an equivalent quantity of the diluted solution of acetic acid (4.5). Determine the exact number of ml beforehand by titration of the ammonia solution using bromothymol blue as indicator (4.6). Mix.

6.2.5 . 12,5 ml zinkacetatoploesning ( 4.1 . ) tilsaettes forsigtigt under omroering , opnaaet ved at rotere kolben , der holdes skraatstillet .

6.2.5. Add, with gently mixing by rotating the tilted flask, 12.5 ml of zinc acetate solution (4.1).

6.2.6 . 12,5 ml kaliumhexacyanoferrat(II)oploesning ( 4.2 . ) tilsaettes paa samme maade som for acetatoploesningen .

6.2.6. Add 12.5 ml of potassium hexacyanoferrate (II) solution (4.2) in the same way as for the acetate solution.

6.2.7 . Kolbeindholdets temperatur justeres til 20 * C , og der fyldes op til 200 ml-maerket med 20 * C vand .

6.2.7. Bring the contents of the flask to 20 oC and make up to the 200 ml mark with water at 20 oC.

Note :

Note:

Under alle trin , der er beskrevet indtil nu , skal tilsaetning af vand eller reagenser foretages paa en saadan maade , at det undgaas at faa dannet luftbobler , og med samme sigte for oeje boer alle blandinger udfoeres ved rotation af kolben fremfor ved omrystning . Hvis der findes luftbobler i kolben , foer der fyldes op til 200 ml-maerket , kan man bidrage til at faa dem fjernet ved kortvarigt at forbinde kolben til en vacuumpumpe under rotation af kolben .

During any of the stages so far described all additions of water or reagents should have been made in such manner as to avoid the formation of air bubbles, and with the same object in view, all mixing should have been carried out by rotation of the flask rather than by shaking. If air bubbles are found to be present before making up to 200 ml volume, their removal can be assisted by temporarily connecting the flask to a vacuum pump, and rotating the flask.

6.2.8 . Kolben lukkes med en toer prop , og der blandes omhyggeligt ved kraftig rystning .

6.2.8. Close the flask with a dry stopper and mix thoroughly by vigorous shaking.

6.2.9 . Blandingen henstilles i faa minutter , og der filtreres derefter gennem et toert filtrerpapir , idet de foerste 25 ml filtrat bortkastes .

6.2.9. Allow to stand for a few minutes and then filter through a dry filter paper, rejecting the first 25 ml of filtrate.

6.2.10 . Direkte polarisation : Bestem den optiske drejning af filtratet ved 20 mere aller mindre 1 * C .

6.2.10. Direct polarization: determine the optical rotation of the filtrate at 20 oC ± 1 oC.

6.2.11 . Inversion : 40 ml af det ovenfor opnaaede filtrat afpipetteres i en 50 ml maalekolbe . 6,0 ml 6,35 M1 saltsyre ( 4.3 . ) eller 7,6 ml 5,00 M1 saltsyre ( 4.3 . ) tilsaettes .

6.2.11. Inversion: pipette 40 ml of the filtrate obtained above into a 50 ml volumetric flask. Add 6,0 ml of 6,35 M hydrochloric acid or 7,5 ml of 5,0 M hydrochloric acid (4.3).

Kolben anbringes paa vandbad ved 60 * C i 15 min . , idet det sikres , at hele kolbens indhold er neddyppet . Der blandes ved at foretage cirkulerende bevaegelser i de foerste 5 min . , inden for hvilken tid kolben boer antage badets temperatur . Der afkoeles til 20 * C og fyldes op til maerket med vand ved 20 * C . Der blandes , og blandingen henstilles i 1 time ved denne temperatur .

Place the flask in a waterbath of 60 oC for 15 minutes, ensuring that the entire bulb of the flask has been immersed. Mix by a rotatory movement during the first five minutes, in which time the contents of the flask should have attained the temperature of the bath. Cool to 20 oC, and make up to volume with water at 20 oC. Mix and allow to stand for one hour at this temperature.

6.2.12 . Invert polarisation

6.2.12. Invert polarization

Drejningen af den inverterede oploesning bestemmes ved 20 mere aller mindre 0,2 * C . ( Hvis temperatur T af vaesken , der skal polariseres , afviger mere end 0,2 * C fra 20 * C under maalingen , maa den temperaturkorrektion , der refereres til under 7.2 . , anvendes ) .

Determine the rotation of the inverted solution at 20 oC ± 0.2 oC. (However, if temperature T of the liquid in the polarization tube differs by more than 0.2 oC during the measurement, the temperature correction referred to under 7.2 must be applied.)

7 . PRAESENTATION AF RESULTATERNE

7. EXPRESSION OF RESULTS

7.1 . Beregningsmetode

7.1. Method of calculation

Saccharoseindholdet beregnes ved hjaelp af foelgende formler :

Calculate the sucrose content by means of the following formulae:

1 . v = M/100 ( 1,08 F + 1,55 P )

(1) v = M1001,08F + 1,55P

2 . S = ( D - 1,25 I )/Q gange V - v/V gange V/L gange M %

(2) ×

hvor :

S = saccharoseindholdet ,

M = vaegten i g af afvejet proeve ,

where:

F = procent fedt i proeven ,

S = sucrose content;

P = procent protein ( N gange 6,38 ) i proeven ,

M = mass of the weighed sample in grams;

V = rumfanget i ml , som proeven er fortyndet op til inden filtreringen ,

F = percentage of fat in the sample;

v = korrektion i ml for rumfanget af bundfaldet , der dannes under klaringen ,

P = percentage Of protein (N x 6.38) in the sample;

D = direkte polarimeterafkoelning ( polarisering foer inversion ) ,

V = volume in ml to which the sample is diluted before filtration;

I = polarimeterafloesning efter inversion ,

v = correction in ml for the volume of the precipitate formed during clarification;

L = laengden i dm af polarimeterroeret ,

D = direct polarimeter reading (polarization before inversion);

Q = inversionsfaktor , hvis vaerdi er angivet nedenfor .

I = polarimeter reading after inversion;

Bemaerkninger :

L = length in dm of the polarimeter tube;

a ) Naar der afvejes noejagtig 40,00 g kondenseret maelk og der anvendes et polarimeter med natriumlys , vinkelgrader og et 2 dm polarimeterroer ved 20,0 mere aller mindre 0,1 * C , kan saccharoseindholdet i normalt kondenseret maelk ( C = 9 ) beregnes ud fra foelgende formel :

Q = inversion factor, the values of which are given below.

S = ( D - 1,25 I ) ( 2,833 - 0,00612 F - 0,00878 P )

Remarks:

b ) Hvis inverspolarisationen maales ved en anden temperatur end 20 * C , skal tallene multipliceres med :

(a) When exactly 40,00 g of condensed milk are weighed and a polarimeter with sodium light, angular degrees and a 2dm polarimeter tube at 20,0 oC ± 0,1 oC is used the sucrose content of normal condensed milk (C = 9) can be calculated from the following formula:

( 1 + 0,0037 ( T - 20 ) ) .

S = (D — 1,25 I) x (2,833 — 0,00612 F — 0,00878 P)

7.2 . Vaerdier for inversionsfaktoren Q

(b) If the invert polarization is measured at a temperature other than 20 oC, the figures should be multiplied by:

De foelgende formler giver noejagtige vaerdier for Q for forskellige lyskilder og indeholder korrektioner for koncentration og temperatur :

(1 + 0,0037 (T — 20).

Natriumlys og polarimeter med vinkelgrader :

7.2. Values of the inversion factor Q

Q = 0,8825 + 0,0006 ( C - 9 ) - 0,0033 ( T - 20 )

The following formulae give accurate values for Q, for various sources of light with corrections for concentration and temperature:

Groent kviksoelvlys og polarimeter med vinkelgrader :

Sodium light and polarimeter with angular degrees:

Q = 1,0392 + 0,0007 ( C - 9 ) - 0,0095 ( T - 20 )

Q = 0,8825 + 0,0006 (C — 9) — 0,0033 (T — 20).

Hvidt lys med dichromatfilter og saccharimeter med internationale sukkerskalagrader :

Mercury green light and polarimeter with angular degrees:

Q = 2,549 + 0,0017 ( C - 9 ) - 0,0095 ( T - 20 ) .

Q = 1,0392 + 0,0007 (C — 9) — 0,0039 (T — 20).

I de ovenfor anfoerte formler er :

White light with dichromate filter and saccharimeter with international sugar scale degrees:

C = procent total sukker i de inverterede proever , som de polariseres .

Q = 2,549 + 0,0017 (C — 9) — 0,0095 (T — 20).

T = Temperaturen af den inverterede oploesning , naar den polariseres .

In the above formulae:

Note 1 :

C = Percentage of total sugars in the inverted solution as polarized,

Det procentmaessige indhold af totalt sukker , C , i den inverterede oploesning kan beregnes ud fra den direkte aflaesning og aendringen ved inversionen paa den saedvanlige maade , idet de normale vaerdier for saccharose , lactose og invertsukkers specifikke rotation anvendes .

T = Temperature of the inverted solution in the polarimetric reading.

Korrektionen 0,0006 ( C - 9 ) osv . er kun noejagtig , naar C er ca . 9 ; for normalt kondenseret maelk kan denne korrektion negligeres , idet C er naesten 9 .

Note 1:

Note 2 :

The percentage of total sugars C in the inverted solution may be calculated from the direct reading and the change on inversion in the usual manner, using the usual values for the specific rotations of sucrose, lactose and invert sugar.

Variationen i temperaturen fra 20 * C paa 1 * C goer ikke nogen stor forskel ved den direkte aflaesning , men en variation paa over 0,2 * C ved den inverte aflaesning noedvendiggoer en korrektion . Korrektionen - 0,0033 ( T - 20 ) osv . er kun noejagtig mellem 18 og 22 * C .

The correction 0,0006 (C — 9) etc., is only accurate when C is approximately 9; for normal condensed milk, this correction can be neglected, C being close to 9.

7.3 . Repetitionsevne

Note 2:

Variation in temperature from 20 oC of 1 oC makes little difference in the direct reading, but variation of over 0,2 oC in the invert reading necessitates a correction. The correction - 0,0033 (T — 20) etc., is only accurate between 18 oC and 22 oC.

METODE 6 : BESTEMMELSE AF MAELKESYRE - OG LACTATINDHOLD

7.3. Repeatability

1 . OMFANG OG ANVENDELSESOMRAADE

The difference between results of two determinations carried out simultaneously or in rapid succession on the same sample, by the same analyst, under the same conditions, shall not exceed 0,3 g of sucrose per 100 g of condensed milk.

Denne metode anvendes ved bestemmelse af indholdet af maelkesyre og lactater udtrykt som maelkesyre i :

METHOD 6: DETERMINATION OF LACTIC ACID AND LACTATES CONTENT

- maelkepulver med hoejt fedtindhold ,

1. SCOPE AND FIELD OF APPLICATION

- maelkepulver eller soedmaelkspulver ,

This method determines the lactic acid and lactates, expressed as lactic acid, contents of:

- pulver af delvis skummet maelk ,

- dried high fat milk or high fat milk powder,

- skummetmaelkspulver .

- dried whole milk or whole milk powder,

2 . DEFINITION

- dried partly skimmed milk or partly skimmed-milk powder,

Indholdet af maelkesyre og lactater i maelkepulvere : indholdet af maelkesyre og lactater udtrykt som maelkesyre som det bestemmes ved den specificerede metode .

- dried skimmed milk or skimmed-milk powder.

3 . PRINCIP

2. DEFINITION

Fedt , protein og lactose fjernes samtidigt fra en oploesning af proeven ved tilsaetning af kobbersulfat og calciumhydroxid efterfulgt af filtrering .

Lactic acid and lactates content of dried milks: the lactic acid and lactates, expressed as lactic acid, contents as determined by the method specified.

Maelkesyren og lactaterne i filtratet omdannes til acetaldehyd med koncentreret svovlsyre under tilstedevaerelse af kobbersulfat .

3. PRINCIPLE

Maelkesyreindholdet bestemmes kolorimetrisk ved anvendelse af p-hydroxydiphenyl .

Fat, protein and lactose are simultaneously removed from a solution of the sample by addition of copper sulphate and calcium hydroxide followed by filtration.

Indholdet af maelkesyre og lactater udtrykkes som mg maelkesyre pr . 100 g fedtfrit toerstof .

The lactic acid and lactates in the filtrate are converted into acetaldehyde by concentrated sulphuric acid in the presence of copper II sulphate.

4 . REAGENSER

The lactic acid content is determined colorimetrically using p-hydroxydiphenyl.

4.1 . Kobber(II)sulfatoploesning : 250 g kobber(II)sulfat ( CuSO4 * 5HO2O ) oploeses i vand , og der fortyndes til 1 000 ml med vand .

The lactic acid and lactates content is expressed as mg of lactic acid per 100 g of solids-non-fat.

4.2.1 . Calciumhydroxidsuspension : 300 g calciumhydroxid ( Ca(OH)2 ) formales i en morter med vand , idet der anvendes i alt 900 ml . Suspensionen boer fremstilles umiddelbart foer anvendelsen .

4. REAGENTS

4.2.2 . Calciumhydroxidsuspension : 300 g calciumhydroxid ( Ca(OH)2 ) formales i en morter med vand , idet der anvendes i alt 1 400 ml . Suspensionen boer fremstilles umiddelbart foer anvendelsen .

4.1. Copper (II) sulphate solution: dissolve 250 g of copper (II) sulphate (CuSO4.5H2O) in water and dilute to 1000 ml with water.

4.3 . Svovlsyre-kobber(II)sulfatoploesning : 0,5 ml kobber(II)sulfatoploesning ( 4.1 . ) tilsaettes 300 ml 95,5 - 97,0 % ( m/m ) H2SO4 ) .

4.2. Calcium hydroxide suspension.

4.4 . p-hydroxydiphenyl * ( C6H5C6H4OH)-oploesning : under svag opvarmning og omrystning oploeses 0,75 g p-hydroxydiphenyl i 5 ml af en vandig oploesning af natriumhydroxid , der indeholder 5 g NaOH pr . 100 ml . Der fortyndes til 50 ml med vand i en maalekolbe . Oploesningen opbevares i en brun farvet flaske paa et moerkt og koeligt sted . Anvendes ikke , hvis der forekommer farveaendringer eller uklarhed . Maximal holdbarhed er 72 timer .

4.2.1. Grind 300 g of calcium hydroxide (Ca(OH)2) in a mortar with water, using totally 900 ml. The suspension should be freshly prepared before use.

4.5 . Maelkesyrestandardoploesning : kort foer brugen oploeses 0,1067 g lithiumlactat ( CH3CHOH-COOLi ) i vand , og der fortyndes til 1 000 ml i en maalekolbe . 1 ml af denne oploesning svarer til 0,1 mg maelkesyre .

4.2.2. Calcium hydroxide suspension: grind 300 g of calcium hydroxide (Ca(OH)2) in a mortar with water, using totally 1400 ml. The suspension should be freshly prepared before use.

4.6 . Standard rekonstitueret maelk : paa forhaand analyseres nogle proever af maelkepulver af hoej kvalitet . Til fremstilling af standardkurven udvaelges den proeve , der har det laveste maelkesyreindhold , idet den ikke maa indeholde mere end 30 mg maelkesyre pr . 100 g fedtfrit toerstof . Foelg fremgangsmaaden under 6.2.1 . og 6.2.2 . nedenfor .

4.3. Sulphuric acid — copper (II) sulphate solution: Add to 300 ml of sulphuric acid, 95,9 to 97,0 % (m/m) of H2SO4, 0,5 ml of the copper (II) sulphate solution (4.1).

5 . APPARATUR

4.4. p-hydroxydiphenyl (C6H5C6H4OH) solution: dissolve, by shaking and by heating slightly 0,75 g of p-hydroxydiphenyl in 5 ml of an aqueous solution of sodium hydroxide, containing 5 g of NaOH per 100 ml. Dilute to 50 ml with water in a volumetric flask. Keep the solution in a brown coloured glass bottle in a dark and cool place. Do not use if the colour changes or tubidity occurs. The maximum shelf life is 72 hours.

5.1 . Analysevaegt .

4.5. Lactic acid standard solution: dissolve, shortly before use, 0,1067 g of lithium lactate (CH3 CHOHCOOLi) in water and dilute to 1000 ml in a volumetric flask. 1 ml of this solution corresponds to 0,1 mg of lactic acid.

5.2 . Spektrofotometer , der kan anvendes til aflaesninger ved en boelgelaengde paa 570 mm .

4.6. Standard reconstituted milk: analyse in advance several samples of high quality dried milk. For the preparation of the calibration curve select the sample having the lowest lactic acid content, containing not more than 30 mg of lactic acid per 100 g of solids-non-fat. Follow the operating procedure described under 6.2.1 and 6.2.2 below.

5.3 . Vandbad ved 30 mere aller mindre 2 * C .

5. APPARATUS

5.4 . Morter og stoeder .

5.1. Analytical balance.

5.5 . Filtrerpapir ( Schleicher og Schull 595 , Whatman 1 eller tilsvarende ) .

5.2. Spectrophotometer suitable for readings at a wavelength of 570 nm.

5.6 . Reagensglas , pyrex eller tilsvarende ( dimensioner 25 - 150 mm ) .

5.3. Waterbath at 30 oC ± 2 oC.

Note :

5.4. Mortar and pestle.

Alle glasvarer skal vaere perfekt rengjorte og kan kun beregnes at skulle anvendes til bestemmelse af maelkesyre og lactat . Glasudstyr indeholdende bundfaldrester skylles med koncentreret saltsyre , inden det vaskes af .

5.5. Filter paper (Schleicher and Schull 595, Whatman 1 or equivalent).

6 . FREMGANGSMAADE

5.6. Test tubes, pyrex or equivalent (dimensions 25 x 150 mm).

6.1 . Blindproeve

Note:

Der udfoeres en blindproeve ved at fylde 30 ml vand i en 50 ml maalekolbe ; kolben behandles som beskrevet under 6.2.4 . - 6.2.11 . inklusive . Hvis blindvaerdien maalt mod vand overstiger en vaerdi svarende til 20 mg maelkesyre pr . 100 g fedtfrit toerstof , boer reagenserne undersoeges og de(t ) urene reagens(er ) erstattes . Blindvaerdien udfoeres samtidigt med analyseproevernes gennemfoerelse .

All glassware must be perfectly clean and designated for use solely in this determination. Rinse glassware containing precipitate residues with concentrated hydrochloric acid before washing.

6.2 . Bestemmelse

6. PROCEDURE

Note : Undgaa tilsmudsning med urenheder isaer af spyt og sved .

6.1. Blank test

6.2.1 . Det fedtfrie toerstofindhold , a g , i proeven bestemmes ved at traekke fedtindholdet ( bestemt ved metode 4 ) og vandindholdet ( bestemt ved metode 2 ) fra 100 .

Carry out a blank test by placing 30 ml of water into a 50 ml graduated tube and treating this tube as described under 6.2.4 to 6.2.11 inclusive. If the blank measured against water exceeds an equivalent of 20 mg of lactic acid per 100 g solids-non-fat, the reagents should be checked and the impure reagents or reagent should be replaced. Carry out the blank test at the same time as the analysis of the sample.

6.2.2 . Med en noejagtighed paa 0,1 mg afvejes ( 1000 / ( a - 10 ) ) g proeve . Til denne maengde saettes 100 ml vand , og der blandes omhyggeligt .

6.2. Determination

6.2.3 . 5 ml af oploesningen afpipetteres i et 50 ml maaleglas , og der fortyndes med vand til ca . 30 ml .

Note: Avoid contamination with impurities especially with saliva and sweat.

6.2.4 . Under omrystning tilsaettes langsomt 5 ml kober(II)sulfatoploesning ( 4.1 . ) , og blandingen henstilles i 10 min .

6.2.1. Determine the solids-non-fat content (a) g of the sample by subtracting the fat content (obtained by method 4) and the moisture content (obtained by method 2) from 100.

6.2.5 . Under omrystning tilsaettes langsomt 5 ml calciumhydroxidsuspension ( 4.2.1 . ) , eller 10 ml hydroxidsuspension ( 4.2.2 . ) .

6.2.2. Weigh 1000a-10g of the sample to the nearest 0,1 g. Add this quantity of sample to 100 ml of

6.2.6 . Der fortyndes til 50 ml vand og rystes kraftigt ; henstilling i 10 min . og derefter filtrering . De foerste gennemloeb bortkastes .

6.2.3. Pipette 5 ml of the solution obtained into a 50 ml graduated tube and dilute with water to about 30 ml.

6.2.7 . 1 ml filtrat afpipetteres i et reagensglas ( 5.6 . ) .

6.2.4. Add slowly while shaking, 5 ml of the copper (II) sulphate solution (4.1) and allow to stand for 10 minutes.

6.2.8 . 6,0 ml svovlsyrekobber(II)sulfatoploesning ( 4.3 . ) tilsaettes til reagensglasset ved hjaelp af en burette eller en gradueret pipette . Der blandes .

6.2.5. Add slowly while shaking, 5 ml of the calcium hydroxide suspension (4.2.1) or 10 ml of the calcium hydroxide suspension (4.2.2).

6.2.9 . Opvarmning paa kogende vandbad i 5 min . Afkoeling til stuetemperatur under rindende vand .

6.2.6. Dilute to 50 ml with water, shake vigorously, allow to stand for 10 minutes then filter. Discard the first runnings.

6.2.10 . 2 draaber 4-hydroxidiphenylreagens ( 4.4 . ) tilsaettes , og der rystes kraftigt for at fordele reagenset jaevnt i hele vaesken . Reagensglasset anbringes paa vandbadet ved 30 mere aller mindre 2 * C ; henstand 15 min . med lejlighedsvis omrystning .

6.2.7. Pipette 1 ml of the filtrate into a test tube (5.6).

6.2.11 . Reagensglasset anbringes paa kogende vandbad i 90 sek . Det afkoeles til stuetemperatur under rindende vand .

6.2.8. Add to the tube by means of a burette or graduated pipette 6.0 ml of the sulphuric acid-copper (II) sulphate solution (4.3). Mix.

6.2.12 . Den optiske taethed maales mod blindproeven ( 6.1 . ) ved den boelgelaengde , der er specificeret i 5.2 . , inden der er gaaet 3 timer .

6.2.9. Heat in the boiling water bath for five minutes. Cool to ambient temperature under running water.

6.2.13 . Hvis den optiske taethed er stoerre end det hoejeste punkt paa standardkurven , gentages bestemmelsen med en passende fortynding af filtratet 6.2.2 .

6.2.10. Add two drops of p-hydroxydiphenyl reagent (4.4) and shake vigorously to spread the reagent evenly throughout the liquid. Place the tube in the waterbath at 30 oC ± 2 oC; leave for 15 minutes shaking from time to time.

6.3 . Fremstilling af standardkurve

6.2.11. Place the tube in the boiling waterbath for 90 seconds. Cool to ambient temperature under running water.

6.3.1 . 5 ml rekonstitueret maelk ( 4.6 . ) afpipetteres i fem 50 ml maaleglas . I disse glas afpipetteres 0 , 1 , 2 , 3 , og 4 ml af standardoploesningen ( 4.5 . ) , saaledes at der opnaas et standardomraade svarende til 0 , 20 , 40 , 60 og 80 mg maelkesyre sat til 100 g fedtfrit toerstof i maelkepulveret .

6.2.12. Measure the optical density against the blank test (6.1) within three hours at the wavelength specified under 5.2.

6.3.2 . Fortynd med vand til ca . 30 ml , og proeven behandles som beskrevet under 6.2.4 . til 6.2.11 . inklusive .

6.2.13. If the optical density exceeds that of the highest point of the standard curve, repeat the test using an adequate dilution of the filtrate obtained under 6.2.6.

6.3.3 . De optiske taetheder af standarderne ( 6.3.1 . ) maales mod blindproeven ( 6.1 . ) ved den boelgelaengde , der er specificeret under 5.2 . I et diagram afbildes de optiske taetheder mod maengderne af maelkesyre , der er angivet under 6.3.1 . , dvs . 0 , 20 , 40 , 60 og 80 g mg pr . 100 g fedtfrit toerstof . Den bedst passende rette linje traekkes gennem punkterne , og standardkurven frembringes ved at parallelforskyde denne linje paa en saadan maade , at den kommer til at gaa gennem begyndelsespunktet .

6.3. Preparation of the standard

7 . PRAESENTATION AF RESULTATERNE

6.3.1. Pipette 5 ml of the reconstituted milk (4.6) into five 50 ml graduated tubes. Pipette into these tubes 0, 1, 2, 3 and 4 ml respectively of the standard solution (4.5), so as to obtain a range of standards corresponding to 0, 20, 40, 60 and 80 mg of added lactic acid per 100 g of solids-non-fat, of the dried milk.

7.1 . Beregningsmetode

6.3.2. Dilute with water to about 30 ml and treat as described under 6.2.4 to 6.2.11.

De optiske taetheder , der er maalt under 6.2.12 . eller 6.2.13 . , omsaettes til mg maelkesyre pr . 100 g fedtfrit toerstof i proeven ved hjaelp af standardkurven . Dette resultat multipliceres med fortyndingsfaktoren , hvor filtratet har vaeret fortyndet som under 6.2.13 .

6.3.3. Measure the optical densities of the standards (6.3.1) against the blank test (6.1) at the wavelength specified under 5.2. Plot in a diagram the optical densities against the quantities of lactic acid given under 6.3.1, i.e. 0 mg, 20 mg, 40 mg, 60 mg and 80 mg per 100 g of solids-non-fat. Draw the best fitting straight line through the points and prepare the standard curve by moving this line parallel to itself in such a way that it passes through the origin.

7.2 . Repetitionsevne

7. EXPRESSION OF RESULTS

Forskellen mellem resultaterne for to bestemmelser , der er udfoert samtidigt eller hurtigt efter hinanden af den samme analytiker , der anvender den samme proeve under de samme betingelser , maa ikke overstige 8 mg pr . 100 g fedtfrit toerstof for indhold op til 80 mg . For hoejere vaerdier boer forskellen ikke overstige 10 % af den laveste vaerdi .

7.1. Method of calculation

METODE 7 : BESTEMMELSE AF PHOSPHATASEAKTIVITET ( MODIFICERET SANDERS OG SAGER METODE )

Convert the optical density measured under 6.2.12 or 6.2.13 into mg of lactic acid per 100 g of solids-non-fat in the sample by reference to the standard curve. Multiply this result by the dilution factor where the filtrate has been diluted according to 6.2.13.

1 . OMFANG OG ANVENDELSESOMRAADE

7.2. Repeatability

Denne metode anvendes ved bestemmelse af phosphataseaktiviteten i :

The difference between the results of two determinations carried out simultaneously or in rapid succession on the same sample, by the same analyst, under the same conditions, shall not exceed 8 mg of lactic acid per 100 g of solids-non-fat for contents up to 80 mg. For higher values, this difference may not exceed 10 % of the lowest value.

- maelkepulver med hoejt fedtindhold ,

METHOD 7: DETERMINATION OF PHOSPHATASE ACTIVITY (MODIFIED SANDERS AND SAGER PROCEDURE)

- maelkepulver eller soedmaelkspulver ,

1. SCOPE AND FIELD OF APPLICATION

- pulver af delvis skummet maelk ,

This method describes the determination of phosphatase activity in:

- skummetmaelkspulver .

- dried high fat milk or high fat milk powder,

2 . DEFINITION

- dried whole milk or whole milk powder,

Phosphataseaktiviteten i maelkepulver er et maal for maengden af aktiv alkalisk phosphatase , der findes . Den udtrykkes som den maengde phenol i mg , der frigoeres af 1 ml rekonstitueret maelk som bestemt ved metoden nedenfor .

- dried partly skimmed milk or partly skimmed-milk powder,

3 . PRINCIP

- dried skimmed milk or skimmed-milk powder.

Phosphataseaktiviteten af maelkepulvere bestemmes ved phosphatasens evne til at frigoere phenol fra dinatriumphenylphosphat . Maengden af phenol , der frigoeres under de beskrevne betingelser , bestemmes ved en spektrofotometrisk maaling af farven , der fremkommer med Gibbs' reagens .

2. DEFINITION

4 . REAGENSER

The phosphatase activity of dried milks is a measure of the quantity of active alkaline phosphatase present. It is expressed as the quantity of phenol in μg liberated by 1 ml of reconstituted milk, as determined by the procedure described below.

4.1 . Oploesning A

3. PRINCIPLE

Bariumborathydroxidpuffer : pH 10,6 mere aller mindre 0,1 ved 20 * C .

The phosphatase activity of dried milks is determined by the ability of the phosphatase to liberate the phenol from disodiumphenylphosphate. The quantity of phenol liberated under prescribed conditions is determined by a spectrophotometric measurement of the colour developed with Gibb's reagent.

25,0 g bariumhodroxid ( Ba(OH)2 , 8H2O ) oploeses i vand , og der fortyndes til 500 ml .

4. REAGENTS

11,0 g borsyre ( H3BO3 ) oploeses i vand , og der fortyndes til 500 ml .

4.1. Solution A

De to oploesninger opvarmes til 50 * C og blandes sammen .

Barium borate hydroxide buffer: pH 10,6 ± 0,1 at 20 o C.

Der omrystes og afkoeles til stuetemperatur . pH justeres til 10,6 mere aller mindre 0,1 med bariumhydroxid , derefter filtrering . Oploesningen opbevares til en taet tillukket beholder . Foer anvendelsen fortyndes pufferen med et tilsvarende volumen vand .

Dissolve: 25,0 g of barium hydroxide (Ba(OH)2.8H2O) in water and dilute to 500 ml.

4.2 . Oploesning B

Dissolve: 11,0 g of boric acid (H3BO3) in water and dilute to 500 ml.

Farvefremkaldelsespuffer .

Warm the two solutions to 50 o C and mix.

6,0 g natriummetaborat ( NaBO2 ) ( eller 1i,6 g NaBO2 . 4H2O ) og 20,0 g natriumchlorid ( NaCl ) oploeses i vand , og der fortyndes til 1 000 ml med vand .

Shake and cool the mixture to room temperature.

4.3 . Oploesning C

Adjust the pH to 10,6 ± 0,1 with the barium hydroxide solution and filter.

Puffersubstratoploesning .

Store the solution in a tightly stoppered container.

4.3.1 . 0,5 g dinatriumphenylphosphat ( Na2C6H5PO4 . 2H2O ) oploeses i 4,5 ml af oploesning B ( 4.2 . ) . Der tilsaettes 2 draaber af oploesning E ( 4.5 . ) , og oploesningen henstilles i 30 min . Farven ekstraheres med 2,5 ml butanol ( 4.10 . ) . Ekstraktionen gentages om noedvendigt . Efter adskillelse bortkastes butanolen . Oploesningen kan opbevares i adskillige dage i koeleskab . Farven fremkaldes og ekstraheres en gang mere inden anvendelsen .

Before use, dilute the buffer with an equal quantity of water.

4.3.2 . 1 ml af denne oploesning afpipetteres i en 100 ml maalekolbe , og der fyldes op til maerket med oploesning A . Pufferoploesningen fremstilles umiddelbart foer anvendelsen .

4.2. Solution B:

4.4 . Oploesning D

Colour development buffer.

Bundfaeldningsmiddel .

Dissolve: 6,0 g of sodium metaborate (NaBO2) (or 12,6 g of NaBO2.4H2O) and 20,0 g of sodium chloride (NaCl) in water and dilute to 1000 ml with water.

3,0 g zinksulfat ( ZnSO4 . 7H2O ) og 0,6 g kobber(II)sulfat ( CuSO4 , 5H2O ) oploeses i vand , og der fyldes op til 100 ml med vand .

4.3. Solution C

4.5 . Oploesning E

Buffer substrate solution.

Gibbs' reagens .

4.3.1. Dissolve 0,5 g of disodiumphenylphosphate (Na2C6H5PO4.2H2O) in 4,5 ml of Solution B (4.2). Add 2 drops of Solution E (4.5) and allow to stand 30 minutes. Extract the colour with 2,5 ml butanol (4.10). If necessary, repeat the colour extraction. After separation, discard the butanol. This solution can be kept for several days in a refrigerator. Develop and extract the colour once more before use.

0,040 g 2,6-dibromoquinon-1,2-chloroimid ( 0-C6H2Br2 , NCl ) oploeses i 10 ml 96 % ethanol . Oploesningen opbevares i en moerk glasflaske , der opbevares i koeleskab . Reagenset bortkastes , hvis det er blevet misfarvet .

4.3.2. Pipette 1 ml of this solution into a 100 ml volumetric flask and make up to volume with Solution A. Prepare the buffer solution immediately before use.

4.6 . Farvefortyndingspuffer .

4.4. Solution D

10 ml af oploesning B ( 4.2 . ) , farvefremkaldelsespuffer , fortyndes til 100 ml med vand .

Precipitant.

4.7 . Kobber(II)sulfatoploesning .

Dissolve 3,0 g of zinc sulphate (ZnSO4.7H2O) and 0,6 g of copper (II) sulphate (CUSO4.5H2O) in water and make up to 100 ml with water.

0,05 g kobber(II)sulfat ( CuSO4 , 5H2O ) oploeses i vand , og der fyldes op til 100 ml med vand .

4.5. Solution E

4.8 . Phenolstandardoploesning .

Gibb's reagent.

0,200 mere aller mindre 0,001 g ren phenol oploeses i vand , og der fyldes op til 100 ml i en maalekolbe med vand . Denne oploesning kan opbevares i adskillige maaneder i koeleskab . 10 ml af denne oploesning fortyndes til 100 ml med vand . Denne fortyndede oploesning indeholder 200 mg phenol pr . ml og kan anvendes til at fremstille mere fortyndede oploesninger .

Dissolve 0,040 g of 2,6-dibromoquinone 1,4 — chloroimide (O.C6H2Br2.NCl) in 10 ml of 96 % ethanol. Store the solution in a dark glass bottle kept in a refrigerator. Discard this reagent when it has become discoloured.

4.9 . Kogt destilleret vand .

4.6. Colour dilution buffer

4.10 . n-butanol .

Dilute 10 ml of Solution B (4.2), colour development buffer, to 100 ml with water.

5 . APPARATUR

4.7. Copper sulphate solution

5.1 . Analysevaegt .

Dissolve 0,05 g of copper (II) sulphate (CUSO4.5H2O) in water and make to 100 ml with water.

5.2 . Vandbad , termostatreguleret ved 37 mere aller mindre 1 * C .

4.8. Phenol standard solution

5.3 . Spektrofotometer , der kan anvendes til aflaesninger ved en boelgelaengde paa 610 nm .

Dissolve 0,200 ± 0,001 g of pure phenol in water and make up to 100 ml in a volumetric flask with water. This solution can be stored for several months in a refrigerator. Dilute 10 ml of this solution to 100 ml with water. This diluted solution contains 200 μg of phenol in 1 ml and can be used for preparing more dilute solutions.

5.4 . Filtrerpapir ( Schleicher og Schull 597 , Whatman 42 eller tilsvarende papir ) .

4.9. Boiled distilled water.

5.5 . Vandbad , kogende .

4.10. n-Butanol.

5.6 . Aluminiumsfolie .

5. APPARATUS

6 . FREMGANGSMAADE

5.1. Analytical balance.

Forholdsregler :

5.2. Waterbath, thermostatically controlled at 37 oC ± 1 oC.

1 . Undgaa direkte udsaettelse for sollys .

5.3. Spectrophotometer suitable for readings at a wavelength of 610 nm.

2 . Alt glasudstyr , propper og andet materiale skal vaere perfekt rengjort . Det anbefales , at det skylles og koges med vand eller dampbehandles .

5.4. Filter paper (Schleicher and Schull 597, Whatman 42 or equivalent filter paper).

3 . Undgaa plastmaterialer ( propper f.eks . ) , da de kan indeholde phenoler .

5.5. Waterbath, boiling.

4 . Spyt indeholder phosphatase ; til smudsning med spor af spyt maa derfor omhyggeligt undgaas .

5.6. Aluminium foil.

6.1 . Fremstilling af proeve

6. PROCEDURE

6.1.1 . Med en noejagtighed paa 0,1 g afvejes 10 g proeve , som oploeses i 90 ml vand . Under oploesningen af pulveret maa temperaturen under ingen omstaendigheder overstige 35 * C .

Precautions:

6.2 . Bestemmelse

1. Avoid direct exposure to sunlight.

6.2.1 . Til hver af to reagensglas saettes 1 ml rekonstitueret maelk fremstillet som beskrevet i 6.1.1 .

2. All the glassware, stoppers and removal material should be perfectly clean. It is recommended that they be rinsed and boiled with water or that they be treated with steam.

6.2.2 . Det ene glas opvarmes paa kogende vandbad i 2 min . Reagensglasset og vandbadet ( 5.5 . ) , eller f.eks . et baegerglas , overdaekkes med aliminiumsfolie ( 6.5 . ) for at sikre , at hele glasset bliver opvarmet . Der afkoeles i koldt vand til stuetemperatur . Dette reagensglas anvendes til blindproeven . Ved alle efterfoelgende operationer behandles de to glas identisk .

3. Avoid using plastic materials (stoppers for example) as they may contain phenols.

6.2.3 . 10 ml oploesning C ( 4.3.2 . ) tilsaettes . Blanding og anbringelse paa vandbadet ved 37 * C ( 5.2 . ) .

4. Saliva contains phosphatase; contamination by traces of saliva must therefore be carefully avoided.

6.2.4 . Inkuber i 60 min . i vandbadet , idet der lejlighedsvis omrystes .

6.1. Preparation of the sample

6.2.5 . Glassene overfoeres straks til et kogende vandbad ( 5.5 . ) og de opvarmes i 2 min . ; afkoeling til stuetemperatur i koldt vand .

6.1.1. Weigh, to within 0.1 g, 10 g of the sample and dissolve in 90 ml of water. The temperature for dissolving the powder shall, under no circumstances, exceed 35 oC.

6.2.6 . 1 ml oploesning D ( 4.4 . ) tilsaettes , og der blandes og filtreres gennem et toert filtrerpapir ; den foerste del af filtratet bortkastes , indtil der opnaas en klar vaeske .

6.2. Determination

6.2.7 . 5 ml af hvert filtrat afpippeteres i reagensglas , 5 ml oploesning B ( 4.2 . ) og 0,1 ml oploesning E ( 4.5 . ) tilsaettes . Der blandes .

6.2.1. Introduce in each of two test tubes 1 ml of reconstituted milk prepared as described in 6.1.1.

6.2.8 . Man lader farven fremkalde i 30 min . ved stuetemperatur ikke udsat for direkte sollys .

6.2.2. Heat one of the tubes in boiling water for two minutes. Cover the tube and the waterbath (5.5) or, for example, a beaker with aluminium foil (5.6) to ensure that the entire tube will be heated. Cool in cold water to room temperature. Use this tube for the blank test. For all subsequent operations treat the two tubes identically.

6.2.9 . Den optiske taethed af proeveoploesningen maales mod blindvaerdien ved den boelgelaengde , der er angivet i 5.3 .

6.2.3. Add 10 ml of Solution C (4.3.2). Mix and place the tube in the waterbath at 37 oC (5.2).

6.2.10 . Bestemmelsen gentages , hvis den optiske taethed af oploesningen er over taetheden for standardproeven med 20 mg phenol , fremstillet som beskrevet i 7 .

6.2.4. Incubate for 60 minutes in the waterbath shaking periodically.

Hvis denne graense er overskredet , fortyndes et passende rumfang rekonstitueret maelk som beskrevet i 6.1.1 . med et passende rumfang af denne maelk omhyggeligt kogt som beskrevet i 6.2.2 . for at inaktivere tilstedevaerende phosphatase .

6.2.5. Transfer the tubes immediately to a boiling waterbath (5.5) and heat for two minutes; cool to room temperature in cold water.

7 . FREMSTILLING AF STANDARDKURVE

6.2.6. Add 1 ml of Solution D (4.4), mix and filter through a dry filter paper; discard the first filtrates until a clear liquid is obtained.

7.1 . I fire 100 ml maalekolber afpipetteres 1 , 3 , 5 og 10 ml standardoploesning , fortyndet som beskrevet i 4.8 . og der fyldes op til maerket med vand ; disse oploesninger indeholder 2 , 6 , 10 og 20 mg phenol pr . ml .

6.2.7. Put 5 ml of each filtrate into test tubes, add 5 ml of Solution B (4.2) and 0.1 ml of Solution E (4.5). Mix.

7.2 . 1 ml af hver standardoploesning ( 7.1 . ) afpipetteres i reagensglas for at opnaa en serie af proever indeholdende 0 ( blindvaerdien faas ved afpipettering af 1 ml vand ) , 2 , 6 , 10 og 20 mg phenol .

6.2.8. Allow the colour to develop at room temperature for 30 minutes away from direct sunlight.

7.3 . I hvert reagensglas afpipetteres efter hinanden 1 ml kobber(II)sulfatoploesning ( 4.7 . ) , 5 ml fortyndingspuffer ( 4.6 . ) , 3 ml vand og 0,1 ml oploesning E . Der blandes .

6.2.9. Measure the optical density of the sample solution, against the blank, at the wavelength indicated in 5.3.

7.4 . Reagensglassene henstilles i 30 min . ved stuetemperatur , vaek fra direkte sollys .

6.2.10. Repeat the determination if the optical density of the solution is above that of the standard sample with 20 μg of phenol prepared according to 7.

7.5 . Absorptioner af oploesningerne i hvert reagensglas maales mod blindvaerdien ved den boelgelaengde , der er angivet i 5.3 .

If this limit is exceeded, dilute a suitable volume of reconstituted milk according to 6.1.1 with a suitable volume of this milk carefully boiled as indicated in 6.2.2 to inactivate the phosphatase present.

7.6 . Standardkurven frembringes ved at afbilde absorptionsvaerdierne mod de maengder af phenol i mg , der blev angivet i 7.2 .

7. PREPARATION OF THE STANDARD CURVE

8 . PRAESENTATION AF RESULTATERNE

7.1. Pipette into four 100 ml volumetric flasks, 1, 3, 5 and 10 ml of the standard solution diluted according to 4.8 and make up to volume with water; these dilutions contain respectively 2, 6, 10 and 20 μg of phenol per ml.

8.1 . Beregning

7.2. Pipette 1 ml of water or 1 ml of each standard solution (7.1) into the test tubes in order to obtain a series of samples containing 0 (blank value obtained using the 1 ml of water) — 2 — 6 — 10 and 20 μg of phenol.

8.1.1 . De tal , der er fremkommet under 6.2.9 . , omregnes til mg phenol ved hjaelp af standardkurven .

7.3. Pipette successively into each test tube 1 ml of the solution of copper (II) sulphate (4.7), 5 ml of the colour dilution buffer solution (4.6), 3 ml of water and 0.1 ml of Solution E (4.5). Mix.

8.1.2 . Phosphataseaktiviteten udtrykt som mg phenol pr . ml rekonstitueret maelk beregnes ud fra foelgende formel :

7.4. Leave the test tubes for 30 minutes at room temperature away from direct sunlight.

phosphataseaktivitet = 2,4 gange P ,

7.5. Measure the absorbance of the solutions in each of the tubes, compared to the blank value, at the wavelength indicated in 5.3.

hvor P = phenolmaengden i mg som beskrevet i 8.1.1 .

7.6. Prepare the standard curve by plotting the absorbance values against the quantities of phenol in μg as indicated in 7.2.

8.1.3 . Hvis det som angivet under 6.2.10 . var noedvendigt at fortynde , skal resultatet fra 8.1.2 . multipliceres med fortyndingsfaktoren .

8. EXPRESSION OF THE RESULTS

8.2 . Repetitionsevne

8.1. Calculation

Forskellen mellem resultaterne for to bestemmelser , der er udfoert samtidigt eller hurtigt efter hinanden af den samme analytiker , der anvender den samme proeve under de samme betingelser , maa ikke overstige 2 mg phenol frigjort pr . 1 ml rekonstitueret maelk .

8.1.1. Convert the figures as determined under 6.2.9 to μg of phenol, by reference to the standard curve.

METODE 8 : BESTEMMELSE AF PHOSPHATASEAKTIVITET ( ASCHAFFENBURG OG MULLEN-PROCEDUREN )

8.1.2. Calculate the phosphatase activity expressed as μg of phenol per ml of reconstituted milk according to the following formula:

1 . OMFANG OG ANVENDELSESOMRAADE

Phosphatase activity = 2,4 x P

Denne metode anvendes ved bestemmelse af phosphataseaktivitet i :

where P = the quantity of phenol in μg according to 8.1.1.

- maelkepulver med hoejt fedtindhold ,

8.1.3. If it was necessary to dilute as indicated under 6.2.10 multiply the result obtained in 8.1.2 by the dilution factor.

- maelkepulver eller soedmaelkspulver ,

8.2. Repeatability

- pulver af delvis skummet maelk ,

The difference between the results of two determinations carried out simultaneously or in rapid succession on the same sample, by the same analyst, under the same conditions, shall not exceed 2 μg of phenol liberated by 1 ml of reconstituted milk.

- skummetmaelkspulver .

METHOD 8: DETERMINATION OF PHOSPHATASE ACTIVITY (ASCHAFFENBURG AND MÜLLEN PROCEDURE)

2 . DEFINITION

1. SCOPE AND FIELD OF APPLICATION

Phosphataseaktiviteten i maelkepulvere er et maal for maengden af aktiv alkalisk phosphatase , der findes i produktet . Den udtrykkes som maengden af p-nitrophenol , der frigoeres af 1 ml af den rekonstituerede proeve under de beskrevne betingelser .

This method describes the determination of phosphatase activity in:

3 . PRINCIP

- dried high fat milk or high fat milk powder,

Den rekonstituerede proeve fortyndes med et buffersubstrat ved pH 10,2 og inkuberes ved en temperatur paa 37 * C i 2 timer . Alkalisk phosphatase , der findes i proven , vil under disse omstaendigheder frigoere p-nitrophenol fra det tilsatte dinatrium-p-nitrophenylphosphat . Den frigjorte p-nitrophenol bestemmes ved direkte sammenligning med standardfarvede glasplader i en simpel komparator under anvendelse af reflekteret lys .

- dried whole milk or whole milk powder,

4 . REAGENSER

- dried partly skimmed milk or partly skimmed-milk powder,

4.1 . Natriumcarbonat-bicarbonatpufferoploesning .

- dried skimmed milk or skimmed-milk powder.

3,5 g vandfrit natriumcarbonat og 1,5 g natriumbicarbonat oploeses i vand , og der fortyndes til 1 000 ml i en maalekolbe med vand .

2. DEFINITION

4.2 . Puffersubstrat .

The phosphatase activity of dried milks is a measure of the quantity of active alkaline phosphatase present in the product. It is expressed as the quantity of p-nitrophenol in micrograms liberated by 1 ml of the reconstituted sample, under the conditions described.

1,5 g dinatrium-p-nitrophenylphosphat oploeses i natriumcarbonat-bicarbonatpufferen ( 4.1 . ) , og der fortyndes til 1 000 ml i en maalekolbe med pufferen .

3. PRINCIPLE

Denne oploesning er stabil i en maaned , hvis den opbevares i koeleskab ( * 4 * C ) , men der boer udfoeres en farvekontrolundersoegelse paa saadanne opbevarede oploesninger - jf . 6 , forholdsregel 3 .

The reconstituted sample is diluted with a buffer substrate at pH 10,2 and incubated at a temperature of 37 oC for two hours. Any alkaline phosphatase present in the sample will, under these circumstances, liberate p-nitrophenol from added disodium p-nitrophenyl phosphate. The p-nitrophenol liberated is determined by direct comparison with standard colour glasses in a simple comparator using reflected light.

4.3 . Klaringsoploesninger .

4. REAGENTS

4.3.1 . Zinksulfatoploesning .

4.1. Sodium carbonate-bicarbonate buffer solution.

30 g zinksulfat ( ZnSO4 oploeses i vand , og der fortyndes til 100 ml i en maalekolbe med vand .

Dissolve 3,5 g of anhydrous sodium carbonate and 1,5 g of sodium bicarbonate in water and dilute to 1000 ml in a volumetric flask with water.

4.3.2 . Kaliumhexacyanoferrat(II)trihydrat ( KuF2%CN)6 * 3H2O ) oploeses i vand , og der fortyndes til 100 ml i en maalekolbe med vand .

4.2. Buffer substate.

5 . APPARATUR

Dissolve 1,5 g of disodium p-nitrophenylphosphate in sodium carbonate-bicarbonate buffer (4.1) and dilute to 1000 ml in a volumetric flask with buffer (4.1).

5.1 . Analysevaegt .

This solution is stable if stored in a refrigerator (≤ 4 oC) for one month but a colour control test should be carried out on such stored solutions — see 6, precaution number 3.

5.2 . Vandbad , termostatreguleret ved 37 mere aller mindre 1 * C .

4.3. Clarification solutions.

5.3 . Komparator , med speciel skive indeholdende standardfarvede glas , kalibrerede i mg p-nitrophenol pr . ml maelk og 2 gange 25 mm kuvette .

4.3.1. Zinc sulphate solution.

6 . FREMGANGSMAADE

Dissolve 30,0 g of zinc sulphate (ZnSO4) in water and dilute to 100 ml in a volumetric flask with water.

Forholdsregler :

4.3.2. Potassium hexacyanoferrate (II) solution.

1 . Efter brugen maa reagensglassene toemmes , skylles i vand , vaskes i varmt vand , indeholdende et alkalisk opvaskemiddel , efterfulgt af en omhyggeligt skylning i rent , varmt hanevand . Sluttelig skal de skylles og toerres foer brugen .

Dissolve 17,2 g of potassium hexacyanoferrate (II) trihydrate (K4Fe(CN)6.3H20) and dilute to 100 ml in a volumetric flask with water.

Pipetter skylles omhyggeligt i rent koldt hanevand umiddelbart efter brugen , efterfulgt af skylning i vand ; de toerres foer brugen .

5. APPARATUS

2 . Reagensglaspropperne skylles omhyggeligt i varmt hanevand umiddelbart efter brugen , efterfulgt af kogning i 2 min . i vand .

5.1. Analytical balance.

3 . Puffersubstratoploesningen ( 4.2 . ) boer holde sig stabil i mindst 1 maaned , hvis den opbevares i koeleskab ved 4 * C eller lavere . Ustabilitet bemaerkes ved dannelsen af en gul farve . Medens proeven altid maales mod en kogt produktkontrol , der indeholder den samme puffersubstratoploesning , anbefales det , at oploesningen ikke anvendes , saafremt den giver en farveaflaesning paa over 10 mg , naar den aflaeses i en 25 mm kuvette i komparatoren mod destilleret vand i den anden 25 mm kuvette .

5.2. Waterbath, thermostatically controlled at 37 oC ± 1 oC.

4 . Anvend en separat pipette til hver proeve og undgaa forurening af pipetten med spyt .

5.3. Comparator, with special disc containing standard colour glasses calibrated in μg p-nitrophenol per ml milk, and 2 x 25 mm cells.

5 . Proeven maa ikke udsaettes for direkte sollys .

6. PROCEDURE

6.1 . Fremstilling af proeve .

Precautions:

10 g pulver oploeses i 90 ml vand . Temperaturen maa under oploesningen ikke overstige 35 * C .

1. After use, test tubes must be emptied, rinsed in water, washed in hot water containing an al kaline detergent, followed by thorough rinsing in clean hot tap water. Finally, they must be rinsed in water and dried before use.

6.2 . Bestemmelse

Pipettes must be thoroughly rinsed in clean cold tap water immediately after use, followed by rinsing in water and dried before use.

6.2.1 . 15 ml puffersubstrat ( 4.2 . ) afpipetteres i et rent , toert reagensglas efterfulgt af 2 ml af den rekonstituerede proeve ( 6.1 . ) , der skal undersoeges . Glasset lukkes , og der blandes ved vending ; placering i vandbadet ved 37 * C ( 5.2 . ) .

2. The test tube stoppers must be thoroughly rinsed in hot tap water immediately after use, followed by boiling for two minutes in water.

6.2.2 . Samtidigt placeres i vandbadet et kontrolglas , der indeholder 15 ml puffersubstrat og 2 ml kogt rekonstitueret proeve som den , der undersoeges .

3. The buffer substrate solution (4.2) should remain stable for at least one month if stored in a refrigerator at 4 oC or less. Any instability is denoted by the formation of a yellow colour. Whilst the test is always read against a boiled product control containing the same buffer substrate solution, it is recommended that the solution should not be used if it gives a colour reading in excess of 10 μg when read in a 25 mm cell in the comparator using distilled water in the other 25 mm cell.

6.2.3 . Efter 2 timer fjernes begge reagensglas fra vandbadet , der tilsaettes 0,5 ml zinksulfatbundfaeldningsmiddel ( 4.3.1 . ) , proppen saettes paa igen og der rystes kraftigt , henstilling i 3 min . 0,5 ml kaliumhexacyanoferrat(II)bundfaeldningsmiddel ( 4.3.2 . ) tilsaettes , og der blandes omhyggeligt ; filtrering gennem foldefilterpapir ( 5.4 . ) , det klare filtrat samles i det rene reagensglas .

4. Use a separate pipette for each sample and avoid contaminating the pipette with saliva.

6.2.4 . Filtratet overfoeres til en 25 mm celle og sammenlignes med filtratet fra den kogte kontrolproeve i komparatoren , idet den specielle skive anvendes ( 5.3 . ) .

5. The test must not be exposed to direct sunlight at any time.

7 . PRAESENTATION AF RESULTATERNE

6.1. Preparation of sample

7.1 . Beregning

Dissolve 10 g of the powder in 90 ml of water. The temperature for dissolving the powder must not exceed 35 oC.

De direkte aflaesninger under 6.2.4 . noteres som mg p-nitrophenol pr . ml proeve eller pr . ml rekonstitueret proeve .

6.2. Determination

7.2 . Repetitionsevne

6.2.1. Pipette 15 ml of buffer substrate (4.2) into a clean, dry test tube, followed by 2 ml of the reconstituted sample (6.1) to be tested. Stopper the tube, mix by inversion and place in the 37 oC water bath (5.2).

6.2.2. At the same time, place in the water bath a control tube containing 15 ml of buffer substrate and 2 ml of boiled reconstituted sample similar to that under test.

6.2.3. After two hours remove both tubes from the water bath, add 0,5 ml of zinc sulphate precipitant (4.3.1), replace the stopper, shake vigorously and allow to stand for three minutes. Add 0,5 ml of potassium hexacyanoferrate (II) precipitant (4.3.2), mix thoroughly and filter through the fluted filter paper (5.4) and collect the clear filtrate in the clean test tube.

6.2.4. Transfer the filtrate to a 25 mm cell and compare against the filtrate of the boiled sample control in the comparator using the special disc (5.3).

7. EXPRESSION OF RESULTS

7.1. Calculation

The direct reading obtained under 6.2.4 is recorded as μg p-nitrophenol per ml sample or per ml of reconstituted sample.

7.2. Repeatability

The difference between the results of two determinations carried out simultaneously or in rapid succession on the same sample, by the same analyst, under the same conditions, shall not exceed 2 μg of p-nitrophenol liberated by 1 ml of reconstituted milk.

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