IE
S.I. No. 200/1984 - European Communities (Marketing of Feedingstuffs) Regulations, 1984.
Table of Provisions (19)
Statutory Text (19 Articles)
1. Where, on official inspection the composition of a straight feedingstuff is found to depart from the declared composition in a manner such as to reduce its value, the following tolerances are permitted:—
( a ) for crude protein
— 2 units for declared contents of 20% or more,
— 10% of the declared content for declared contents of less than 20% but not less than 10%,
—1 unit for declared contents of less than 10%;
( b ) for total sugars, reducing sugars, sucrose, lactose, glucose (dextrose) and total solids
— 2 units for declared contents of 20% or more,
— 10% of the declared content for declared contents of less than 20% but not less than 5%.
— 0.5 unit for declared contents of less than 5%;
( c ) for starch and inulin
— 3 units for declared contents for 30% or more,
— 10% of declared content for declared contents of less than 30% but not less than 10%,
— 1 unit for declared contents of less than 10%;
( d ) for crude oils and fats
— 1.8 units for declared contents of 15% or more,
— 12% of the declared contents for declared contents of less than 15% but not less than 5%,
— 0.6 unit for declared contents of less than 5%;
( e ) for moisture
— 1 unit for declared contents of 10% or more,
— 10% of the declared content for declared contents of less than 10% but not less than 5%,
— 0.5 unit for declared contents of less than 5%;
( f ) for total phosphorus, sodium, calcium carbonate, calcium, magnesium, acid index and matter insoluble in light petroleum
— 1.5 units for declared contents (values) of 15% (15) or more, as appropriate,
— 10% of the declared content (value) for declared contents (values) of less than 15% (15) but not less than 2%(2), as appropriate;
— 0.2 unit for declared contents (values) of less than 2% (2) as appropriate;
( g ) for ash insoluble in hydrochloric acid and chlorides expressed as NaCl
— 10% of the declared content for declared contents of 3% or more,
— 0.3 unit for declared contents of less than 3%;
( h ) for carotene, vitamin A and xanthophyll
— 30% of the declared content;
(i) for methionine, cystine, lysine and volatile nitrogenous bases
— 20% of the declared content;
(j) for crude fibre
—2.2 units for declared contents of 11% or more,
— 20% of the declared content for declared contents of less than 11% but not less than 7%,
— 1.4 units for declared contents of less than 7%,
(k) for crude ash
— 1.5 units for declared contents of 10% or more,
— 15% of the declared content for declared contents of less than 10% but not less than 4%,
— 0.6 unit for declared contents of less than 4%;
(l) for Hectolitre Weight
—1.5 units.
(m) for Nitrogen
— 0.4 unit.
2. Where, on official inspection the composition of a compound feedingstuff other than for dogs and cats is found to depart from the declared composition, the following tolerances are permitted:—
(i) Where the content recorded is less than the declared content
(a) Crude protein and skim milk powder
— 2 units for declared contents of 20% or more,
— 10% of the declared content for declared contents of less than 20% but not less than 10%,
— 1 unit for declared contents of less than 10%;
(b) Soluble protein
— 3 units for declared contents of 25% or more,
— 12% of the declared content for declared contents of less than 25% but not less than 15%,
— 1.8 units for declared contents of less than 15%;
(c) Total sugar
— 2 units for declared contents of 20% or more,
—10% of the declared content for declared contents of less than 20% but not less than 10%,
— 1 unit for declared content of less than 10%;
(d) Starch and total sugar plus starch
— 2.5 units for declared contents of 25% or more,
— 10% of the declared content for declared contents of less than 25% but not less than 10%,
— 1 unit for declared contents of less than 10%;
(e) Crude oils and fats
— 1.5 units for declared contents of 15% or more,
— 10% of the declared content for declared contents of less than 15% but not less than 8%,
— 0.8 unit for declared contents of less than 8%;
(f) Sodium and magnesium
— 1.5 units for declared contents of 15% or more,
— 10% of the declared content for declared contents of less than 15% but not less than 7.5%,
— 0.75 unit for declared contents of less than 7.5% but not less than 5%,
— 15% of the declared content for declared contents of less than 5% but not less than 0.7%,
— 0.1 unit for declared contents of less than 0.7%;
(g) Total phosphorus and calcium
— 1.2 units for declared contents of 16% or more,
—7.5% of the declared content for declared contents of less than 16% but not less than 12%,
— 0.9 unit for declared contents of less than 12% but not less than 6%,
— 15% of the declared content for declared contents of less than 6% but not less than 1%,
— 0.15 unit for declared contents of less than 1%;
(h) Methionine and lysine
—15% of the declared content;
(i) Cystine
— 20% of the declared content.
(ii) Where the content recorded is more than the declared content
(a) Moisture
— 1 unit for declared contents of 10% or more,
— 10% of the declared content for declared contents of less than 10% but not less than 5%,
— 0.5 unit for declared contents of less than 5%;
(b) Crude ash
— 1.5 units for declared contents of 10% or more,
— 15% of the declared content for declared contents of less than 10% but not less than 4%,
— 0.6 unit for declared contents of less than 4%;
(c) Crude fibre
— 2.2 units for declared contents of 11% or more,
— 20% of the declared content for declared contents of less than 11% but not less than 7%,
— 1.4 units for declared contents of less than 7%;
(d) Ash insoluble in hydrochloric acid
— 1 unit for declared contents of 10% or more,
— 10% of the declared content for declared contents of less than 10% but not less than 4%,
— 0.4 unit for declared contents of less than 4%;
(iii) Where the variation noted is in the opposite direction to those referred to respectively in paragraphs 2 (i) and 2 (ii)
(a) — crude protein, crude oils and fats, total sugar, starch: tolerance twice that permitted for these substances in paragraph 2 (i);
(b) — total phosphorus, calcium, magnesium, sodium, crude ash, crude fibre; tolerance three times that permitted for these substances in paragraphs 2 (i) and 2 (ii).
3. Where, on official inspection the composition of a compound feedingstuff for dogs and cats is found to depart from the declared composition, the following tolerances are permitted
(i) Where the content recorded is less than the declared content;
(a) Crude protein
— 3.2 units for declared contents of 20% or more;
— 16% of the declared content for declared contents of less than 20% but not less than 12.5%;
— two units for declared contents of less than 12.5%.
(b) Crude oils and fats
— 2.5 units of the declared content.
(ii) Where the content recorded is more than the declared content
(a) Moisture
— 3.0 units for declared contents of 40% or more,
— 7.5% of the declared content for declared contents of less than 40% but not less than 20%,
— 1.5 units for declared contents of less than 20%.
(b) Crude ash
— 1.5 units of the declared content.
(c) Crude Fibre
— 1.0 unit of the declared content.
(iii) Where the variation noted is in the opposite direction to those referred to respectively in paragraphs 3 (i) and 3 (ii)
(a) Crude protein
— tolerance twice that permitted for this substance in paragraph 3 (i).
(b) Crude oils and fats
— identical tolerance,
(c) Crude ash, crude fibre
— tolerance three times that permitted for these substances in paragraph 3 (ii).
GIVEN under my Official Seal, this 31st day of July, 1984.
AUSTIN DEASY,
Minister for Agriculture.
EXPLANATORY NOTE.
The Regulations which cover the marketing of all feedingstuffs other than specified exceptions, lay down quality, labelling and packing requirements for straight and compound feedingstuffs, as well as official methods of sampling and analysis. They revoke the Fertilisers, Feeding Stuffs and Mineral Mixtures Regulations 1957 (
S.I. No. 264 of 1957
) other than Regulation 13, insofar as they relate to feedingstuffs and mineral mixtures.
4. Apparatus
4.1 Water bath.
4.2 Vacuum evaporation apparatus with round flasks of different capacities.
4.3 Glass chromatography tubes (length: approximately 300 mm; internal diameter: approximately 13 mm).
4.4 Spectrophotometer. Measurements in the UV require silica cells.
4.5 UV lamps suitable for 364 nm.
5.
Procedure
5.1 Test sample.
From the finely divided sample, take a test sample proportional to the presumed Vitamin A content, thus
0.1-1.0 g for concentrates (contents greater than 20,000 IU/g);
3.0-5.0 g for premixtures (contents of between 400-20,000 IU/g);
10-20 g for mineral mixtures;
30 g for Group A products.
Immediately place the test sample in a flask with a ground glass stopper.
NB. All operations must be carried out away from direct light, if necessary in brown glass equipment.
5.2 Hydrolysis and extraction
1
Add successively to the test sample 40 ml ethanol (3.1), 2 ml sodium ascorbate solution (3.2)
2
, 10 ml potassium hydroxide solution (3.4) and 2 ml sodium sulphide solution (3.13).
Heat for 30 minutes at 70-80°C under a reflux condenser and then leave to cool under a stream of water. Add 50 ml ethanol (3.1) and 100 ml 1, 2-dichlorethane (3.7). Shake vigorously and then decant the supernatant liquid into a decanting container. Add to the container 150 ml potassium hydroxide solution (3.5), shake for 30 seconds and leave to stand until the layers are separated. Collect the dichlorethane layer (lower layer) in a decanting container, add 40 ml potassium hydroxide solution (3.6), shake for 10 seconds and leave to stand until the layers are separated. Collect the dichlorethane layer in a decanting container and wash 6-8 times with 40 ml lots of water until free of alkali (phenolphthalein test). Collect the dichlorethane layer and remove the last traces of water using strips of filter paper.
Evaporate to dryness an aliquot part of the solution under vacuum and on the water bath at 40°C. Rapidly treat the residue with 5 ml light petroleum (3.8).
For Group A products, chromatograph as shown in 5.3.1.
For Group B products, transfer the solution to a 50 ml graduated flask, make up to volume with light petroleum (3.8), mix and measure the optical density as shown in 5.4.2.
5.3 Chromatography
5.3.1 Group A products.
Fill a chromatography tube (4.3) to a height of 200 mm with aluminium oxide (3.9) previously slurried with light petroleum (3.8). Place in the tube the solution obtained in 5.2 and immediately add 20 ml light petroleum (3.8). Elute successively with 10 ml lots of the light petroleum solutions at 4, 8, 12, 16 and 20 per cent diethyl ether (3.12) under pressure or partial vacuum, the rate of flow being 2-3 drops per second.
The carotene is eluted first
3
. The Vitamin A is generally eluted with the light petroleum solution at 20% diethyl ether (3.12). The elution is followed under UV light (brief irradiation of the column with the mercury lamp). The fluorescent zone of the Vitamin A is clearly separated from the yellow xanthophyll zones following it. Collect the eluate fraction containing the Vitamin A in an Erlenmeyer flask.
1
For milk feeds and products with a tendency to agglomerate or swell double the quantity of the reagents shown in the first and second paragraphs of 5.2.
2
Sodium ascorbate need not be added when hydrolysis is carried out in a notrogen atmosphere.
3
Carotene content may be determined by optical density measurement at 450 nm;
5.3.2 Group B products.
Chromatography must only be carried out if the optical density measurements obtained in 5.4.2 do not conform to the requirements given in 5.4.2.
If chromatography proves necessary, place in the chromatography column an aliquot part of the solution in the light petroleum obtained in 5.2, containing approximately 500 IU of Vitamin A, and chromatography as shown in 5.3.1.
5.4 Measurement of the optical density
5.4.1 Group A products.
Evaporate to dryness under vacuum the eluate containing the Vitamin A obtained in 5.3.1. Treat the residue with 2 ml benzene (3.14). Take 0.3 ml of this solution and add 3 ml of the Carr-Price reagent (3.16). A blue colouring develops. Measure the optical density with the spectrophotometer at 610 nm exactly 30 seconds after the reaction has begun. Determine the Vitamin A content by reference to a standard curve obtained from benzene solutions of increasing Vitamin A — standard concentrations treated with Carr-Price reagent (2-16 IU Vitamin A — standard (3.17) per 0.3 ml benzene (3.14) +3 ml Carr-Price reagent (3.16)). The standard curve must be checked regularly and frequently using the standard and a freshly prepared Carr-Price reagent solution.
5.4.2 Group B products.
Take an aliquot part of the solution in light petroleum obtained in 5.2 containing approximately 200 IU Vitamin A. Evaporate to dryness under vacuum and treat the residue with 25 ml isopropanol (3.18). Measure the optical density in the spectrophotometer at 325, 310 and 334 nm. The absorption maximum is located at 325 nm. The Vitamin A content of the solution is calculated as follows
E
325
18.30=IU of Vitamin A/ml
However, the ratio of the optical densities
E
310
: E
325
and E
334
: E
325
must be 6 : 7=0.857.
If one of these ratios differs appreciably from this value (< 0.830 or > 0.880), the measurement of the optical densities must be preceded by chromatography in accordance with the method given in 5.3.2. If the measurement of the optical densities carried out after chromatography shows that the above mentioned ratios still differ appreciably from the value of 0.857 (< 0.830 or > 0.880), the determination must be carried out in accordance with the method given for Group A products.
6.
Calculation of results
Calculate the Vitamin A content of the sample taking into account the weight of the test sample and the dilutions carried out in the course of analysis. Express the results in IU of Vitamin A per kg of feeding stuff, concentrate or premixture.
7.
Repeatability
The difference between the results of two parallel determinations carried out on the same sample must not exceed
20%, relative to the higher result, for Vitamin A contents lower than 75,000 IU/kg;
15,000 IU for contents between 75,000–150,000 IU/kg;
10%, relative to the higher result, for contents between 150,000–250,000 IU/kg;
25,000 IU for contents between 250,000–500,000 IU/kg;
5%, relative to the higher result, for contents greater than 500,000 IU/kg.
THIRD SCHEDULE
PART II
(Regulations 11 (5) and 17)
CERTIFICATE OF RESULT OF ANALYSIS
Laboratory Ref. No. ..........................................................
Sample of ............................................................ ..................................................
Marked .....................................
received by the State Chemist on ............................................................ ............................................................ ..........
from ............................................................ ............................................................ ............................................................
taken at the premises of ............................................................ ............................................................ ..........................
Method of analysis used where
two or more methods are applicable ............................................................ ............................................................ .......
This is to certify that the above-mentioned sample, which was duly fastened and sealed, has been analysed under the direction of the State Chemist and that the result of the analysis is as follows:—
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This certificate is given under the European Communities (Marketing of Feedingstuffs) Regulations 1984.
Date ...................................
Signed ............................................................ ............................................................ ...
State Chemist
FOURTH SCHEDULE
LIMITS OF ERROR REFERRED TO IN REGULATION 18(2).
5. PROTEINS.
5.1 DETERMINATION OF CRUDE PROTEIN.
1.
Purpose and scope
To determine the crude protein content of feedingstuffs on the basis of the nitrogen content, determined according to the Kjeldahl method.
2.
Principle
The sample is digested by mineral acid. The acid solution is alkalized with a sodium hydroxide solution. The ammonia released is removed by distillation and collected in a measured quantity of sulphuric acid, the excess of which is titrated with a solution of sodium hydroxide.
3.
Reagents
3.1 Potassium sulphate.
3.2 Catalyst: cupric oxide CuO or crystallized cupric sulphate CuSO
4
.5H
2
O or mercury or mercuric oxide HgO.
3.3 Granulated zinc.
3.4 Sulphuric acid, d : 1.84.
3.5 Sulphuric acid, 0.1 N.
3.6 Sulphuric acid, 0.5 N.
3.7 Methyl red indicator: dissolve 300 mg of methyl red in 100 ml of 95-96% (v/v) ethanol.
3.8 Sodium hydroxide solution, 40% (w/v).
3.9 Sodium hydroxide solution, 0.1 N.
3.10 Sodium hydroxide solution, 0.25 N.
3.11 Sodium sulphide, saturated solution.
3.12 Sodium thiosulphate solution, 8% (w/v) Na
2
S
2
O
3
.5H
2
O.
3.13 Granulated pumice stone, washed in hydrochloric acid and ashed.
4.
Apparatus
Apparatus for digestion by combustion and for distillation by the Kjeldahl method (see item 7.1).
5.
Procedure
5.1 Digestion
Weigh, to the nearest mg. approximately 1 g of sample and place in the flask of the digestion apparatus. Add 10 g of potassium sulphate (3.1), and appropriate quantity of catalyst (3.2) (0.3-0.4 g of cupric oxide or 0.9-1.2 g of cupric sulphate or a drop of mercury or 0.6-0.7 g of mercuric oxide), 25 ml of sulphuric acid (3.4) and a few granules of pumice stone (3. 13). Mix. Heat the flask moderately at first, shaking from time to time, until the mass has carbonized and the foam has disappeared; then heat more intensively until the liquid is boiling steadily. Prevent the sides from becoming overheated and organic particles from sticking to them. When the solution becomes clear and colourless (or light green if a copper-based catalyst is used), continue to boil for another hour, then leave to cool.
5.2 Distillation
Carefully add 250-350 ml of water, stirring all the while to dissolve the sulphates completely; leave to cool. Add a few granules of zinc (3.3).
Place in the collecting flask of the distillation apparatus and exactly measured quantity of 25 ml of sulphuric acid 0.1 N (3.5) or 0.5 N (3.6) depending on the presumed nitrogen content (see item 7.2), and add a few drops of methyl red indicator (3.7).
Connect the distillation flask to the condenser of the distillation apparatus and immerse the end of the condenser in the liquid contained in the collecting flask to a depth of at least 1 cm (see item 7.3). Slowly pour 100 ml of 40% sodium hydroxide solution (3.8) into the distillation flask through the dropping funnel. If a mercury-based catalyst has been used, also add either 10 ml of sodium sulphide solution (3.11), or 25 ml of sodium thiosulphate solution (3.12).
Heat the flask in such a way that approximately 150 ml of liquid is distilled in 30 minutes. At the end of this time, check the pH of the resulting distillate with litmus paper. If the reaction is alkaline, continue distillation. Discontinue when the distillate becomes neutral to litmus paper. During distillation keep the colouration under observation and shake the contents of the collecting flask from time to time. If the liquid turns yellow, immediately add an exactly measured volume of sulphuric acid 0.1 N (3.5) or 0.5 N (3.6).
5.3 Titration
In the collecting flask titrate the excess sulphuric acid with sodium hydroxide solution 0.1 N (3.9) or 0.25 N (3.10), depending on the normality of the sulphuric acid used, until the colour turns pale yellow.
5.4 Verification of the method
To establish whether the reagents are free from nitrogen, carry out a blank test (distillation and titration) omitting the sample to be analysed. To check that the apparatus is working properly and that the correct application of the method is used carry out the analysis (digestion, distillation and titration) on 1.5-2.0 g of acetanilide (m.p. 114°C : %N : 10.36) in the presence of 1 g of nitrogen-free sucrose; 1 g of acetanilide corresponds to 14.80 ml of sulphuric acid 0.5 N.
6.
Calculation of results
Determine the volume of sulphuric acid neutralised. 1 ml of sulphuric acid 0.1 N corresponds to 1.4 mg of nitrogen. Multiply the quantity of nitrogen by the factor 6.25. Express the result as a percentage of the sample.
7.
Repeatability
The difference between the results of two parallel determinations carried out on the same sample must not exceed
— 0.2%, in absolute value, for crude protein contents of less than 20% ;
— 1.0%, relative to the higher result, for contents of not less than 20% and not more than 40%;
— 0.4%, in absolute value, for contents of more than 40%.
8.
Observations
8.1 Certain apparatus requiring transference between digestion and distilation may be used. If such apparatus is used, the transfer must be carried out without loss.
8.2 For products with a low nitrogen content, the volume of sulphuric acid 0.I N to be placed in the collecting flask may be reduced, if necessary, to 10 or 15 ml and made up to 25 ml with water.
8.3 If the flask of the distillation apparatus is not fitted with a dropping funnel, add the sodium hydroxide immediately before connecting the flask to the condenser, pouring the liquid slowly down the sides of the condenser so that it does not mix with the acid solution.
5.2 DETERMINATION OF CRUDE PROTEIN DISSOLVED BY PEPSIN AND HYDROCHLORIC ACID.
1.
Purpose and scope
To determine the fraction of crude protein dissolved by pepsin and hydrochloric acid under defined conditions. It is applicable to all feedingstuffs.
2.
Principle
The sample is heated for 48 hours at 40°C in a solution of pepsin hydrochloride. The suspension is filtered and the nitrogen content of the filtrate determined according to the method for the determination of crude protein.
3.
Reagents
3.1 Hydrochloric acid, d : 1.125.
3.2 Hydrochloric acid, 0.075 N.
3.3 2.0 U/mg pepsin; pepsin activity is defined in the method for the Estimation of Pepsin Activity specified in these Regulations, and must be established according to that method.
3.4 About 0.2% (w/v) freshly prepared solution of pepsin in hydrochloric acid (3.2); activity: 400 U/litre.
3.5 Anti-foaming emulsion (e.g. silicone).
3.6 All reagents listed under 3 in the method for the determination of crude protein. (Method 5.1).
4.
Apparatus
4.1 Water bath or incubator, set at 40°C ± 1°C.
4.2 Kjeldahl digestion and distillation apparatus.
5.
Procedure
5.1 Preparation of solution (see item 7.2).
Weigh, to the nearest mg, approximately 2.g of the sample and place in a 500 ml graduated flask. Add 450 ml of pepsin hydrochloride solution (3.4) previously heated to 40°C and shake to prevent the formation of agglomerates. Check that the pH of the suspension is less than 1.7. Place the flask in the water bath or incubator (4.1) and leave there for 48 hours. Shake after 8, 24 and 32 hours. After 48 hours, add 15 ml of hydrochloric acid (3.1), cool to 20°C, make up to volume with water and filter.
5.2 Digestion
Take 250 ml of the filtrate and place in the flask of the distillation apparatus (4.2). Add the reagents necessary for digestion indicated in the second sentence of 5.1 of the method for the determination of crude protein. Mix and bring to the boil. If any foam should form, add a few drops of anti-foaming emulsion (3.5). Continue to boil vigorously until the water has been almost completely evaporated. Reduce the heat and carefully eliminate the last traces of water. When the solution becomes clear and colourless (or light green if a copper-based catalyst is used), continue to boil for another hour. Leave to cool.
5.3 Distillation and titration
Proceed as indicated in 5.2 and 5.3 of the method for the determination of crude protein (Method 5.1).
5.4 Blank test
Carry out a blank test applying the same procedure but omitting the sample to be analysed.
6.
Calculation of results
Subtract the volume of sulphuric acid neutralised in the blank test from that neutralised by the test sample. 1 ml of sulphuric acid 0.1 N corresponds to 1.4 mg of nitrogen.
Multiply the quantity of nitrogen by the factor 6.25. Express the result as a percentage of the sample.
7.
Repeatability
The difference between the results of two parallel determinations carried out on the same sample must not exceed
— 0.4%, in absolute value, for contents of less than 20%;
— 2.0%, relative to the higher result, for contents of not less than 20% and not more than 40%.
—0.8%, in absolute value, for contents of more than 40%.
8.
Observations
8.1 The values obtained by this method have no direct connection with digestibility in vivo.
8.2 Products with an oil or fat content exceeding 10% must first be defatted by extraction with petroleum ether (boiling point 40-60°C).
5.3 ESTIMATION OF PEPSIN ACTIVITY.
1.
Purpose and scope
To establish the activity of the pepsin used in the determination of crude protein dissolved by pepsin and hydrochloric acid.
2.
Principle
Haemoglobin is treated with pepsin in a hydrochloric acid medium under defined conditions. The non-hydrolysed fraction of the protein is precipitated in trichloroacetic acid. Sodium hydroxide and Folin-Ciocalteu reagent are added to the filtrate. The optical density of this solution is measured at 750 nm and the corresponding quantity of tyrosine is read from a calibration curve.
Definition: The unit of pepsin is defined as being the quantity of that enzyme which, under the conditions of the method, liberates per minute, a quantity of hydroxyaryl groups which, when stained with the Folin-Ciocalteu reagent, has an optical density corresponding to that of one µmole of tyrosine stained in the same manner.
3.
Reagents
3.1 Hydrochloric acid, 0.2 N.
3.2 Hydrochloric acid, 0.06 N.
3.3 Hydrochloric acid, 0,025 N.
3.4 Trichloroacetic acid, solution, 5% (w/v).
3.5 Sodium hydroxide solution 0.5 N.
3.6 Folin-Ciocalteu reagent: place 100 g of sodium tungstate (Na
2
WO
4
.2H
2
O), 25 g of sodium molybate (Na
2
Mo0
4
.2H
2
O) and 700 ml of water in a 2 litre round-bottomed flask fitted with a standard ground-glass joint. Add 50 ml of phosphoric acid (d : 1.71) and 100 ml of concentrated hydrochloric acid (d : 1.19), connect a reflux condenser to the flask, bring to the boil and keep the solution gently boiling for 10 hours. Leave to cool, detach the reflux condenser, add 175 g of lithium sulphate (Li
2
SO
4
.2H
2
O), 50 ml of water and 1 ml of bromine. Boil for 15 minutes to eliminate excess bromine. Leave to cool, transfer the solution to a 1 litre graduated flask, make up to volume with water, homogenize and filter. No greenish colouration must remain. Before use, dilute 1 volume of the reagent with 2 volumes of water.
3.7 Haemoglobin solution: weigh a quantity of haemoglobin (approx. 2 g of protein substratum determined according to Anson) corresponding to 354 mg of nitrogen
1
and place in a 200 ml flask fitted with a standard ground-glass joint. Add a few ml of hydrochloric acid (3.2), connect the flask to the vacuum pump and shake until the haemoglobin has completely dissolved. Release the vacuum and, while shaking, add hydrochloric acid (3.2) to make up to 100 ml. Prepare immediately before use.
3.8 Standard tyrosine solution: dissolve 181.2 mg of tyrosine in the hydrochloric acid (3.1) and make up to 1 litre with the same acid (stock solution). Take 20.0 ml and dilute to 100 ml with hydrochloric acid (3.1). 1 ml of this solution contains 0.2 µmole of tyrosine.
4.
Apparatus
4.1 Water bath set at 25°C ± 0.1°C.
4.2 Spectrophotometer.
4.3 Chronometer, accuracy: 1 second.
4.4 pH-meter.
5.
Procedure
5.1 Preparation of the solution (see item 7.1 ).
Dissolve 150 mg of pepsin in 100 ml of hydrochloric acid (3.2). Pipette 2 ml of the solution into a 50 ml graduated flask and make up to volume with hydrochloric acid (3.3). The pH, checked with the pH-meter, must be 1.6±0.1. Immerse the flask in the water bath (4.1).
5.2 Hydrolysis.
Pipette 5.0 ml of haemoglobin solution (3.7) into a test tube, heat to 25°C in the water bath (4.1), add 1.0 ml of the pepsin solution obtained in 5.1 and mix with a glass rod thickened at one end, with about 10 back-and-forth movements. Leave the test tube in the water bath at 25°C for exactly 10 minutes, timed from the addition of the pepsin solution (duration and temperature must be strictly observed). Then add 10.0 ml of trichloroacetic acid solution (3.4) previously heated to 25°C, mix and filter through a dry filter.
1
Determine the nitrogen content by the semi-micro Kjeldahl method (theoretical content: 17.7% of nitrogen).
5.3 Development of colouration and measurement of optical density.
Pipette 5.0 ml of the filtrate into a 50 ml Erlenmeyer flask. Add 10.0 ml of sodium hydroxide solution (3.5) and, shaking constantly, 3.0 ml of dilute Folin-Ciocalteu reagent (3.6). After 5-10 minutes, determine the optical density of the solution with the spectrophotometer at 750 nm in cells 1 cm thick against water.
5.4 Blank test
For each determination, carry out a blank test as follows
Pipette 5.0 ml of haemoglobin solution (3.7) into a test tube, heat to 25°C in the water bath (4.1), add 10.0 ml of trichloroacetic acid solution (3.4) previously heated to 25°C, mix, then add 1.0 ml of the pepsin solution obtained in 5.1. Mix with a glass rod and leave the test tube in the water bath (4.1) at 25°C for exactly 10 minutes. Mix and filter through a dry filter. Follow the procedure indicated in 5.3.
5.5 Calibration curve.
Place 1.0, 2.0, 3.0, 4.0 and 5.0 ml aliquots of standard tyrosine solution (3.8), corresponding to 0.2, 0.4, 0.6, 0.8 and 1.0 vmoles of tyrosine respectively in 50 ml Erlenmeyer flasks. Complete the series with a reference solution free from tyrosine. Make up the volumes to 5.0 ml with hydrochloric acid (3.1). Add 10.0 ml of sodium hydroxide solution (3.5) and, shaking constantly, 3.0 ml of dilute Folin-Ciocalteu reagent (3.6). Measure the optical density as indicated in the last sentence of 5.3. Trace the calibration curve by plotting the optical densities against the quantities of tyrosine.
6.
Calculation of results
From the calibration curve read the quantity of tyrosine, in vmoles, corresponding to the optical density of the coloured solution, corrected on the basis of the blank value.
The pepsin activity, in vmoles, of tyrosine at 25°C, per mg and per minute, is calculated by using the formula
Units per mg(U/mg)=
where
a = quantity of tyrosine, in vmoles, read from the calibration curve;
p = weight in mg of the quantity of pepsin added in 5.2.
7.
Observations
7.1 The quantity of pepsin to be dissolved must be such that, on final photometric measurement, an optical density of 0.35±0.035 is obtained.
7.2 Two units per mg obtained by this method correspond to: 3.64 Anson milliunits/mg (vmoles of tyrosine/mg/min at 35.5°C) or 36,400 commercial units/g (vmoles of tyrosine/g in 10 min at 35.5°C).
5.4 DETERMINATION OF MILK POWDER.
1.
Purpose and scope
To determine the skim milk powder content of milk replacers. Large quantities of buttermilk and/or of certain non-milk proteins may lead to interferences.
2.
Principle
The casein is extracted with sodium citrate solution and precipitated as para-casein by rennet, following adjustment of the calcium ion concentration. The nitrogen content of the para-casein is determined according to the method for the determination of crude protein (see method 5.1). The skim milk powder content is calculated on the basis of a minimum casein content of 27.5%.
3.
Reagents
3.1 Trisodium citrate, dihydrate (1% w/v solution).
3.2 Calcium chloride (saturated aqueous solution at 20°C). Dissolve about 90 g of anhydrous calcium chloride in 100 ml of distilled water by warming and slightly stirring. Leave overnight at 20°C (if a deposit of crystals is not formed, add more calcium chloride and repeat the procedure), filter the liquid next day and store the filtered solution in a well closed bottle at 20°C.
3.3 0.1 N sodium hydroxide.
3.4 0.1 N hydrochloric acid.
3.5 Liquid calf rennet (Standard strength of 1 : 10,000). Store in a refrigerator at 4 to 6°C.
3.6 All reagents listed under 3 in method 5.1 determination of crude protein.
4.
Apparatus
4.1 Centrifuge (2,000 to 3,000 rpm).
4.2 Magnetic stirrer.
4.3 Thermostatically-controlled water bath at 37°C.
4.4 pH meter.
4.5 Kjeldahl digestion and distillation apparatus.
5.
Procedure
5.1 Dissolution of the casein
Weigh, to the nearest 2 mg, 1 g of sample directly into a 50 ml centrifuge tube, add 30 ml of anaqueous solution of trisodium citrate (3.1) previously heated to 45°C. Mix with the aid of the magnetic stirrer (4.2) for at least five minutes. Centrifuge at 500 g (2,000 to 3,000 rpm) for 10 minutes and decant the clear aqueous supernatant into a 150 to 200 ml beaker. Ensure that the sediment remains in the centifuge tube. Carry out two further extractions on the residue, according to the same procedure, and combine the extracts. If a layer of oil forms at the surface, cool in a refrigerator until the fat solidifies and remove the solid layer with a spatula.
5.2 Coagulation of casein with the enzymes of rennet.
While stirring continuously, add dropwise 1 ml of a saturated solution of calcium chloride (3.2) to the total aqueous extract (about 100 ml). Adjust the pH to 6.4-6.5 with solutions of NaOH (3.3) or HCl (3.4) Place in the thermostatically controlled water bath at 37°C for 15 to 20 minutes to obtain saline balance. This becomes more evident by the formation of a light turbidity. (If a precipitate forms it must be removed by further centrifugation at 1,000 rpm for five minutes and the supernatant is decanted without washing the sediment). Remove the extract from the water bath and immediately add 0.5 ml of liquid rennet (3.5) dropwise, with constant stirring. Coagulation appears in one to two minutes. Leave at a temperature of 20 to 37°C for 10 to 15 minutes and break the coagulum by stirring. Transfer the casein quantitatively on to a fast ashless filter while washing three times with 15 ml of distilled water (3 x 15 ml). Filtration must be completed within two hours.
5.3 Determination of casein nitrogen
The filter paper and precipitate is placed in a Kjeldahl flask and nitrogen is determined by the Kjeldahl method as described in method 5.1 determination of crude protein.
5.4 Blank test
Carry out a blank test applying the same procedure but omitting the sample to be analysed.
6.
Calculation of results
Subtract the volume of sulphuric acid neutralised in the blank test from that neutralised by the test sample. 1 ml of sulphuric acid (0. 1 N) corresponds to 1.4 mg of nitrogen. The percentage of skimmed-milk powder in the compound feedingstuff is calculated by the following formula
where N is the percentage of para-casein nitrogen; 27.5 is the factor for converting determined casein into a percentage of skimmed-milk powder.
7.
Repeatability
1.0% relative to the higher result.
5.5 DETERMINATION OF VOLATILE NITROGENOUS BASES
5.5.1
Microdiffusion Method
1.
Purpose and scope
To determine the content of volatile nitrogeneous bases, expressed as ammonia in feedingstuffs.
2.
Principle
The sample is extracted with water and the solution clarified and filtered. The volatile nitrogenous bases are displaced by microdiffusion using a solution of potassium carbonate, collected in a solution of boric acid and titrated with sulphuric acid.
3.
Reagents
3.1 Trichloroacetic acid solution 20% (w/v).
3.2 Indicator: dissolve 33 mg of bromocresol green and 65 mg of methyl red in 100 ml of 95%-96% (v/v) of ethanol.
3.3 boric acid solution: in a 1 litre graduated flask dissolve 10 g of boric acid in 200 ml of 95%-96% (v/v) ethanol and 700 ml of water. Add 10 ml of indicator (3.2). Mix and, if necessary, adjust the colour of the solution to light red by adding a solution of sodium hydroxide. 1 ml of this solution will fix a maximum of 300 vg of ammonia.
3.4 Potassium carbonate saturated solution: dissolve 100 g of potassium carbonate in 100 ml of boiling water. Cool and filter.
3.5 Sulphuric acid 0.02 N.
4.
Apparatus
4.1 Mixer (tumbler): approximately 35 to 40 rpm.
4.2 Glass or plastic Conway cells (see diagram).
4.3 Microburettes graduated in 1/100 ml.
5.
Procedure
Weigh to the nearest mg, approximately 10 g of sample and place in a 200 ml graduated flask. Add 100 ml water and mix in the tumbler (4.1) for 30 mins. Add 50 ml of trichloroacetic acid solution (3.1), make up to volume with water, shake vigorously and filter. If the ammonia content of the sample exceeds 0.6% dilute an aliquot before proceeding to the next stage.
Using a pipette, introduce 1 ml of boric acid solution (3.3) into the central part of the Conway cell and 1 ml of the sample filtrate into the crown of the cell. Cover partially with the greased lid. Drop 1 ml of saturated potassium carbonate solution (3.4) quickly into the crown and close the lid so that the cell is airtight. Turn the cell carefully, rotating it in a horizontal plane so that the two reagents are mixed. Leave to incubate either for at least four hours at room temperature or for one hour at 40°C.
Using a microburette (4.3), titrate the volatile bases in the boric acid solution with sulphuric acid 0.02 N (3.5).
Carry out a blank test using the same procedure but without a sample to be analysed.
6.
Calculation of results
1 ml of sulphuric acid 0.02 N corresponds to 0.34 mg of ammonia.
Express the result as a percentage of the sample.
7.
Repeatability
The difference between the results of two parallel determinations carried out on the same sample should not exceed
10% relative to the higher value, for ammonia contents less than 1.0%.
0.1% in absolute value for ammonia contents equal to or greater than 1.0%.
5.5 DETERMINATION OF VOLATILE NITROGENOUS BASES
5.5.2
Distillation method
1.
Purpose and scope
To determine the content of volatile nitrogenous bases, expressed as ammonia, in fish-meal containing practically no urea. It is applicable only to ammonia contents of less than 0.25%.
2.
Principle
The sample is extracted with water and the solution clarified and filtered. The volatile nitrogenous bases are displaced at boiling point by adding magnesium oxide and collected in a specific quantity of sulphuric acid, the excess of which is back-titrated with a solution of sodium hydroxide.
3.
Reagents
3.1 Trichloroacetic acid solution 20% (w/v).
3.2 Magnesium oxide.
3.3 Anti-foaming emulsion (e.g. silicone).
3.4 Sulphuric acid 0.1 N.
3.5 Sodium hydroxide solution 0.1 N.
3.6 Methyl red indicator; dissolve 300 mg of methyl red in 100 ml of 95-96% (v/v) ethanol.
4.
Apparatus
4.1 Mixer (tumbler): approximately 35 to 40 rpm.
4.2 Distilling apparatus of the Kjeldahl type.
5.
Procedure
Weigh to the nearest mg, approximately 10 g of sample and place it in a 200 ml graduated flask. Add 100 ml of water and mix in the tumbler (4.1) for 30 mins. Add 50 ml of trichloroacetic acid solution (3.1), make up to volume with water, shake vigorously and filter through a pleated filter.
Take a quantity of clear filtrate appropriate for the presumed content of volatile nitrogenous bases (100 ml is usually suitable). Dilute to 200 ml and add 2 g of magnesium oxide (3.2) and a few drops of anti-foaming emulsion (3.3). The solution should be alkaline to litmus paper; otherwise add some magnesium oxide (3.2) Distil about 150 ml of the solution in the Kjeldahl apparatus and collect the distillate in an Erlenmeyer flask containing an accurately measured volume (25 to 50 ml) of sulphuric acid 0.1 N (3.4). While distilling, avoid overheating of the sides. Boil the sulphuric acid solution for two minutes, cool it and back titrate the excess sulphuric acid with the sodium hydroxide solution 0.1 N (3.5) in the presence of the methyl red indicator (3.6).
Carry out a blank test using the same procedure but without a sample to be analysed.
6.
Calculation of results
1 ml of sulphuric acid 0.1 N corresponds to 1.7 mg of ammonia.
7.
Repeatability
The difference between the results of two parallel determinations carried out on the same sample should not exceed, in relative value, 10% of ammonia.
6. MINERAL SUBSTANCES.
6.1 DETERMINATION OF CRUDE ASH.
1.
Purpose and scope
To determine the crude ash content of feeding stuffs.
2.
Principle
The sample is ashed at 550°C; the residue is weighed.
3.
Reagent
Ammonium nitrate solution, 20% (w/v).
4.
Apparatus
4.1 Hot plate.
4.2 Electric muffle-furnace with thermostat.
4.3 Crucibles for ashing made of platinum or an alloy of platinum and gold, or silica or porcelain.
5.
Procedure
Weigh, to the nearest mg, approximately 5 g of the sample (2.5 g in the case of products which have a tendency to swell) and place in a crucible for ashing (4.3) which has first been heated to 550°C ± 5°C, cooled in a "desiccator" and weighed. Place the crucible on the hot plate (4.1) and heat gradually until the substance carbonises. Put the crucible into the muffle-furnace (4.2) set at 550°C ± 5°C. Keep at this temperature until white, light grey or reddish ash is obtained which appears to be free from carbonaceous particles. Place the crucible in a desiccator, leave to cool and weigh immediately.
6.
Calculation of results
Calculate the weight of the residue and express the result as a percentage of the sample.
7.
Observations
7.1 Substances which are difficult to ash must be subjected to an initial ashing of at least 3 hours, cooled and then a few drops of 20% solution of ammonium nitrate (3) added to it (carefully, to avoid dispersal of the ash or the formation of lumps). Continue ashing after drying in the oven. Repeat the operation as necessary until ashing is complete.
7.2 In the case of substances resistant to the treatment described under 7.1, proceed as follows: after ashing for 3 hours, place the ash in warm water and filter through a small, ash-free filter. Ash the filter and its contents in the original crucible. Place the filtrate in the cooled crucible, evaporate until dry, ash and weigh.
7.3 In the case of oils and fats, weigh accurately a sample of approximately 25 g in a suitably sized crucible. Carbonise by setting light to the substance with a strip of ash-free filter paper. After combustion, moisten with as little water as possible. Dry and ash as described under 5.
6.2 DETERMINATION OF ASH WHICH IS INSOLUBLE IN HYDROCHLORIC ACID.
1.
Purpose and scope
To determine the content, in feeding stuffs, of mineral substances which are insoluble in hydrochloric acid. Ether of two methods can be used, depending on the nature of the sample.
1.1 Method A: applicable to straight organic feeding stuffs and to compound feedingstuffs other than those specified under Method B.
1.2 Method B: applicable to mineral compounds and mixtures and to compounds feeding stuffs whose content of substances insoluble in hydrochloric acid, as determined by Method A, is greater than 1%.
2.
Principle
2.1
Method A: the sample is ashed, the ash boiled in hydrochloric acid and the insoluble residue filtered and weighed.
2.2 Method B: the sample is treated with hydrochloric acid. The solution is filtered, the residue ashed and the ash thus obtained treated in accordance with Method A.
3.
Reagents
3.1 Hydrochloric acid solution, 3 N.
3.2 Trichloroacetic acid solution, 20% (w/v).
3.3 Trichloroacetic acid solution, 1% (w/v).
4.
Apparatus
4.1 Hot plate.
4.2 Electric muffle-furnace with thermostat.
4.3 Crucibles for ashing made of platinum or an alloy of platinum and gold, or of silica or porcelain.
5.
Procedure
5.1 Method A
Ash the sample using the method prescribed in this schedule for the determination of crude ash. Ash obtained from that analysis may also be used.
Place the ash in a 250-400 ml beaker using 75 ml of hydrochloric acid 3 N (3.1). Bring slowly to the boil and boil gently for 15 minutes. Filter the warm solution through an ash-free filter paper and wash the residue with warm water until the acid reaction is no longer visible. Dry the filter containing the residue and ash in a tared crucible at a temperature of not less than 550°C and not more than 700°C. Cool in a desiccator and weigh.
5.2 Method B
Weigh, to the nearest mg, approximately 5 g of the sample, and place in a 250-400 ml beaker. Add 25 ml of water and 25 ml of hydrochloric acid 3 N (3.1) successively, mix and wait for effervescence to cease. Add a further 50 ml of hydrochloric acid 3 N (3.1). Wait for any release of gas to cease then place the beaker in a boiling water bath and keep it there for 30 minutes (or longer if necessary) in order to hydrolyse thoroughly any starch which may be present.
Filter while warm through an ash-free filter and wash the filter in 50 ml of warm water (see observation, 7). Place the filter containing the residue in a crucible for ashing (4.3), dry and ash at a temperature of not less than 550°C and not more than 700°C. Place the ash in a 250-400 ml beaker using 75 ml of hydrochloric acid 3 N (3.1); continue as described in the second sub-paragraph of 5.1.
6.
Calculation of results
Calculate the weight of the residue and express the result as a percentage of the sample.
7.
Observation
If filtration proves difficult recommence the analysis, replace the 50 ml of hydrochloric acid 3 N (3.1) by 50 ml of 20% trichloroacetic acid (3.2) and washing the filter in a warm solution of 1% trichloroacetic acid (3.3).
6.3 DETERMINATION OF WATER-SOLUBLE CHLORIDES.
1.
Purpose and scope
To determine the content of water-soluble chlorides, expressed as sodium chlorides, in all feeding stuffs and mineral mixtures.
2.
Principle
The chlorides are dissolved in water. If the product contains organic matter it is clarified. The solution is slightly acidified with nitric acid and the chlorides precipitated in the form of silver chloride by means of a solution of silver nitrate. The excess silver nitrate is titrated with a solution of ammonium thiocyanate, by Volhard's method.
3.
Reagents
3.1 Ammonium thiocyanate solution, 0.1 N.
3.2 Silver nitrate solution, 0.1 N.
3.3 Ammonium ferric sulphate, saturated solution.
3.4 Nitric acid, d : 1.38.
3.5 Diethyl ether or light petroleum, boiling range 40-60°C.
3.6 Acetone.
3.7 Carrez I solution: dissolve in water 21.9 g of zinc acetate (Zn CH
3
COO)
2
.2H
2
O) and 3 g of glacial acetic acid. Make up to 100 ml with water.
3.8 Carrez II solution: dissolve in water 10.6 g of potassium ferrocyanide, K
4
[Fe(CN)
6
].3H
2
O. Make up to 100 ml with water.
3.9 Active carbon, free from chlorides and not absorbing them.
4.
Apparatus
Mixer (tumbler): approximately 35-40 rpm.
5.
Procedure
5.1 Preparation of the solution.
According to the nature of the sample, prepare a solution as shown under 5.1.1, 5.1.2 or 5.1.3.
At the same time carry out a blank test omitting the sample to be analysed.
5.1.1 Samples free from organic matter.
Weigh, to the nearest mg, a sample of not more than 10 g and containing not more than 3 g of chlorine in the form of chlorides. Place with 400 ml of water in a 500 ml volumetric flask at approximately 20°C. Mix for 30 minutes in the tumbler, bring up to volume, mix and filter.
5.1.2 Samples containing organic matter, excluding the products listed under 5.1.3.
Weigh, to the nearest mg, approximately 5 g of the sample and place with 1 g of active carbon (3.9) in a 500 ml volumetric flask. Add 400 ml of water at approximately 20°C and 5 ml of Carrez solution I (3.7), stir, then add 5 ml of Carrez solution II (3.8). Mix for 30 minutes in the tumbler, bring up to volume, mix and filter.
5.1.3 Cooked feeding stuffs, linseed cakes and meal, products rich in linseed meal and other products rich in mucilage or in colloidal substances (for example, dextrinated starch).
Prepare the solution as described under 5.1.2 but do not filter. Decant (if necessary centrifuge), remove 100 ml of the supernatant liquid and transfer to a 200 ml measuring flask. Mix with acetone (3.6) and bring up to volume with this solvent, mix and filter.
5.2 Titration
Using a pipette, transfer to an Erlenmeyer flask from 25-100 ml of the filtrate (according to the assumed chlorine content) obtained as described under 5.1.1, 5.1.2 or 5.1.3. The aliquot portion must not contain more than 150 mg of chlorine (Cl). Dilute if necessary to not less than 50 ml with water, add 5 ml of nitric acid (3.4), 20 ml of saturated solution of ammonium ferric sulphate (3.3) and two drops of ammonium thiocyanate solution (3.1) transferred by means of a burette filled up to the zero mark. Using a burette, transfer the silver nitrate solution (3.2) in such a way that an excess of 5 ml is obtained. Add 5 ml of diethyl ether or light petroleum (3.5) and shake hard to coagulate the precipitate.
Titrate the excess silver nitrate with the ammonium thiocyanate solution (3.1) until the reddish-brown tint has lasted for 1 minute.
6.
Calculation of results
.
The amount of chlorine (W), expressed as sodium chloride, present in the volume of filtrate taken for titration is calculated by using the following formula
W=5.845(V
1
-V
2
) mg
where
V
1
=ml of silver nitrate solution 0.1 N added;
V
2
=ml of ammonium thiocyanate solution 0.1 N used for titration.
If the blank test indicates that silver nitrate solution 0.1 N has been consumed deduct this value from the volume (V
1
-V
2
).
7.
Observations
7.1 Titration may also be carried out by potentiometry.
7.2 In the case of products which are very rich in oils, first defat with light petroleum.
7.3 In the case of fish meal, titration may be carried out by Mohr's method.
6.4 DETERMINATION OF CARBONATES.
1.
Purpose and scope
To determine the content of carbonates, conventionally expressed as calcium carbonate, in most feedingstuffs.
However in certain cases (for example, with iron carbonate) a special method must be used.
2.
Principle
The carbonates are decomposed in hydrochloric acid; the carbon dioxide released is collected in a graduated tube and its volume compared with that released under the same conditions by a known quantity of calcium carbonate.
3.
Reagents
3.1 Hydrochloric acid, d : 1.10.
3.2 Calcium carbonate.
3.3 Sulphuric acid, approximately 0.1 N, coloured with methyl red.
4.
Apparatus
Scheibler-Deitrich apparatus (see diagram fig. 1) or equivalent apparatus.
5.
Procedure
According to the sample's carbonate content, weigh a portion of the sample as shown below
0.5 g for products containing from 50-100 per cent of carbonates, expressed as calcium carbonate;
1 g for products containing from 40-50 per cent of carbonates, expressed as calcium carbonate;
2-3 g for other products.
Place the portion of the sample in the special flask (4) of the apparatus, fitted with a small tube of unbreakable material containing 10 ml of hydrochloric acid (3.1), and connect the flask to the apparatus. Turn the three-way cock (5) so that the tube (1) connects with the outside. Using the mobile tube (2) which is filled with coloured sulphuric acid (3.3) and connected to the graduated tube (1), bring the level of the liquid up to the zero mark. Turn the cock (5) in order to connect up tubes (1) and (3) and check that the level is at zero.
Run the hydrochloric acid (3.1) slowly over the portion of the sample, tilting the flask (4). Make the pressure equal by lowering the tube (2). Shake the flask (4) until the release of carbon dioxide has stopped completely.
Restore pressure by bringing the liquid back to the same level in tubes (1) and (2). After a few minutes, when the volume of gas has become constant, take the reading.
Carry out a control test in the same conditions on 0.5 g of calcium carbonate (3.2).
6.
Calculation of results
The content in grams of carbonates, expressed as calcium carbonate, as a percentage of the sample, is calculated by using the formula
where
V=ml of CO
2
released by the portion of the sample;
T=ml of CO
2
released by 0.5 g of CaCO
3
;
W=weight, in grams, of the portion of the sample.
7.
Observations
7.1 When the portion of the sample weighs more than 2 g first place 15 ml of water in the flask (4) and mix before beginning the test. Use the same volume of water for the control test.
7.2 If the apparatus used has a different volume from that of the Schiebler-Dietrich apparatus, the portions taken from the sample and from the control substance and the calculation of the results must be adapted accordingly.
6.5 DETERMINATION OF CALCIUM.
1.
Purpose and scope
To determine the total calcium content of feeding stuffs.
2.
Principle
The sample is ashed, the ash treated with hydrochloric acid and the calcium precipitated as calcium oxalate. The precipitate is dissolved in sulphuric acid and the oxalic acid formed is titrated with a solution of potassium permanganate.
3.
Reagents
3.1 Hydrochloric acid, d : 1.14.
3.2 Nitric acid, d : 1.40.
3.3 Sulphuric acid, d : 1.13.
3.4 Ammonia, d : 0.98.
3.5 Cold saturated solution of ammonium oxalate.
3.6 Citric acid solution. 30% (w/v).
3.7 Ammonium chloride solution, 5% (w/v).
SCHEIBLER—DIETRICH APPARATUS FOR DETERMINATION OF CO
2
Scale
1
/8
(measured in mm)
Fig. 1.
3.8 Bromocresol green solution, 0.04% (w/v).
3.9 Potassium permanganate solution, 0.1 N.
4.
Apparatus
4.1 Electric muffle-furnace with air circulation and thermostat.
4.2 Platinum, silica or porcelain crucibles for ashing.
4.3 Glass filter crucibles of G4 porosity.
5.
Procedure
Weigh, to the nearest mg, approximately 5 g of the sample (or more if necessary), incinerate at 550C and transfer the ash to a 250 ml beaker.
Add 40 ml of hydrochloric acid (3.1), 60 ml of water and a few drops of nitric acid (3.2). Bring to the boil and keep at boiling point for 30 minutes. Cool and transfer the solution to a 250 ml volumetric flask. Rinse, bring the volume up to the mark with water, mix and filter.
Using a pipette, transfer to a 250 ml beaker an aliquot containing 10-40 mg of calcium according to the assumed calcium content. Add 1 ml of citric acid solution (3.6) and 5 ml of ammonium chloride solution (3.7).
Make the volume up to approximately 100 ml with water. Bring to the boil, add eight to ten drops of bromocresol green solution (3.8) and 30 ml of a warm solution of ammonium oxalate (3.5). If a precipitate forms, dissolve it by adding a few drops of hydrochloric acid (3.1).
Neutralise very slowly with ammonia (3.4), stirring continuously until a pH reading of 4.44.6 is obtained (i.e. when the indicator changes colour). Place the beaker in a boiling water bath and keep there for 30 minutes to allow the precipitate which has formed to settle. Remove the beaker from the water bath. Leave it to stand for an hour and filter through a G4 filter crucible (4.3).
Wash the beaker and the crucible with water until the excess ammonium oxalate is completely removed (the absence of chloride in the washing water indicates that they have been sufficiently washed).
Dissolve the precipitate on the filter in 50 ml of warm sulphuric acid (3.3). Rinse the crucible with warm water and make the filtrate up to approximately 100 ml. Bring the temperature up to 70-80C and titrate drop by drop with a solution of potassium permanganate (3.9) until a pink colour is obtained which lasts for 1 minute.
6.
Calculation of results
1 ml of potassium permanganate 0.1 N corresponds to 2.004 mg of calcium. Express the result obtained as a percentage of the sample.
7.
Observations
7.1 For very low contents of calcium proceed as follows: filter the calcium oxalate precipitate through an ash-free filter paper. After washing, dry the filter and ash at 550C in a platinum crucible. Redissolve the residue in a few drops of sulphuric acid (3.3), evaporate until dry, incinerate again at 550C and weigh. If W is the weight of the calcium sulphate obtained, the calcium content of the aliquot amount taken as a sample=W.0.2944.
7.2 If the sample consists solely of mineral substances, dissolve in hydrochloric acid (3.1) without ashing it first. In the case of products such as calcium aluminium phosphate which are difficult to dissolve in acid, melt as follows by an alkaline process before dissolving: mix the sample to be analysed thoroughly in a platinum crucible with a mixture five times its weight, consisting of equal amounts of potassium carbonate and sodium carbonate. Heat carefully until the mixture is completely melted. Cool and dissolve in hydrochloric acid.
7.3 If the magnesium content of the sample is high, precipitate the calcium oxalate a second time.
6.6 DETERMINATION OF MAGNESIUM.
6.6.1
Atomic absorption method.
1.
Purpose and scope
To determine the magnesium content of feeding stuffs. It is particularly appropriate for determining magnesium contents lower than 5%.
2.
Principle
The sample is ashed and dissolved in dilute hydrochloric acid. If it contains no organic substances, it is dissolved directly in dilute hydrochloric acid. The solution is diluted and the magnesium content determined by atomic absorption spectrophotometry at 285.2 nm, by comparison with standard solutions.
3.
Reagents
3.1 Hydrochloric acid, d : 1.16.
3.2 Hydrochloric acid, d : 1.19.
3.3 Magnesium ribbon or wire, or magnesium sulphate heptahydrate, dried at room temperature.
3.4 Strontium salt solution (chloride or nitrate) at 2.5% (w/v) strontium (=76.08 g SrCl
2
.6H
2
O or 60.38 g Sr(NO
3
)2 per 1000 ml).
3.5 Standard magnesium solution: weigh, to the nearest mg, 1 g magnesium (3.3) which has previously had its oxide coating carefully removed, or the corresponding quantity (10.143 g) of magnesium sulphate heptahydrate (3.3). Place in a 1000 ml graduated flask, add 80 ml hydrochloric acid (3.1), leave to dissolve and make up to 1000 ml with water. 1 ml of this solution contains 1.000 mg magnesium.
4.
Apparatus
4.1 Platinum, silica or porcelain ashing crucibles.
4.2 Thermostatically controlled electric muffle-furnace.
4.3 Atomic absorption spectrophotometer.
5.
Procedure
5.1 Preparation of the sample solution.
5.1.1 Feeding stuffs composed exclusively of mineral substances.
Weigh, to the nearest mg, approximately 5 g of the sample into a 500 ml graduated flask with 250-300 ml water. Add 40 ml hydrochloric acid (3.1), bring to the boil and keep the liquid gently boiling for 30 minutes. Leave to cool, make up to volume with water, mix and filter into a dry beaker through a dry pleated filter. Discard the first 30 ml of the filtrate. In the presence of silica, treat 5 g of sample with a sufficient quantity (15-30 ml) of hydrochloric acid (3.2), evaporate to dryness on a water bath and transfer to an oven at 105C for 1 hour. Proceed as from the third sentence of 5.1.2.
5.1.2 Feeding stuffs composed predominantly of mineral substances.
Weigh, to the nearest mg, approximately 5 g of the sample into a crucible (4.1) and ash at 550C in the muffle-furnace (4.2) until an ash which is free from carbonaceous particles is obtained, and leave to cool. In order to eliminate silica, add to the ash a sufficient quantity (15-30 ml) of hydrochloric acid (3.2), evaporate to dryness on a water bath and transfer to an oven at 105°C for 1 hour. Treat the residue with 10 ml hydrochloric acid (3.1) and transfer to a 500 ml graduated flask using warm water. Leave to cool and make up to volume with water. Mix and filter into a dry beaker through a dry pleated filter. Discard the first 30 ml of the filtrate.
5.1.3 Feeding stuffs composed predominantly of organic substances.
Weigh, to the nearest mg, approximately 5 g of the sample into a crucible (4.1) and ash at 550C in the muffle-furnace (4.2) until an ash which is free from carbonaceous particles is obtained. Treat the ash with 5 ml hydrochloric acid (3.2), evaporate to dryness on a water bath and then dry for 1 hour in the oven at 105C in order to render the silica insoluble. Treat the ash with 5 ml hydrochloric acid (3.1), transfer to a 250 ml graduated flask using warm water, bring to the boil, leave to cool and make up to volume with water. Mix and filter into a dry beaker through a dry pleated filter. Discard the first 30 ml of the filtrate.
5.2. Measurement by atomic absorption
By diluting the standard solution (3.5) with water, prepare at least 5 reference solutions of increasing concentration, corresponding to the optimal measuring range of the spectrophotometer (4.3). Add to each solution 10 ml strontium salt solution (3.4) and then make up the volume to 100 ml with water. Dilute with water one aliquot of the filtrate obtained from 5.1.1, 5.1.2 or 5.1.3, so as to obtain a magnesium concentration which is within the limits of concentration of the reference solutions. The hydrochloric acid concentration of this solution must not exceed 0.4 N. Add 10 ml strontium salt solution (3.4) and then make up the volume to 100 ml with water. Measure the absorption of the solution to be determined and of the reference solutions at 285.2 nm.
6.
Calculation of results
Calculate the quantity of magnesium in the sample by relation to the reference solutions. Express the result as a percentage of the sample.
7.
Repeatability
The difference between the results of two parallel determinations carried out on the same sample must not exceed 5%, relative to the higher result.
6.7 DETERMINATION OF TOTAL PHOSPHORUS.
6.7.1
Spectrophotometric method.
1.
Purpose and scope
To determine the total phosphorus content of feeding stuffs and mineral mixtures low in phosphorus.
2.
Principle
The sample is mineralised, either by dry combustion (in the case of organic feeding stuffs) or by acid digestion (in the case of mineral compounds and liquid feeding stuffs), and placed in an acid solution. The solution is treated with molybdovanadate reagent. The optical density of the yellow solution thus formed is measured in a spectrophotometer at 430 nm.
3.
Reagents
3.1 Calcium carbonate.
3.2 Hydrochloric acid d : 1.1 (approximately 6 N).
3.3 Nitric acid, d : 1.045.
3.4 Nitric acid, d : 1.38-1.42.
3.5 Sulphuric acid, d : 1.84.
3.6 Molybdovanadate reagent: mix 200 ml of ammonium heptamolybdate solution (3.6.1), 200 ml of ammonium monovanadate solution (3.6.2) and 134 ml of nitric acid (3.4) in a 1 litre graduated flask. Make up to volume with water.
3.6.1 Ammonium heptamolybdate solution: dissolve in hot water 100 g of ammonium heptamolybdate (NH
4
)
6
Mo
7
O
24
4H
2
O. Add 10 ml of ammonia (d : 0.91) and make up to 1 litre with water.
3.6.2 Ammonium monovanadate solution: dissolve 2.35 g of ammonium monovanadate (NH
4
VO
3
) in 400 ml of hot water. Stirring constantly, slowly add 20 ml of dilute nitric acid (7 ml of HNO
3
(3.4) +13 ml of H
2
O) and make up to 1 litre with water.
3.7 Standard solution of 1 mg phosphorus per ml: dissolve 4.387 g of potassium dihydrogen phosphate (KH
2
PO
4
) in water. Make up to 1 litre with water.
4.
Apparatus
4.1 Silica or porcelain ashing crucibles.
4.2 Electric muffle-furnace with thermostat set at 550C.
4.3 Kjeldahl flask.
4.4 Graduated flasks and precision pipettes.
4.5 Spectrophotometer.
4.6 Test tubes with stoppers.
5.
Procedure
5.1 Preparation of the solution.
According to the nature of the sample, prepare a solution as indicated in 5.1.1 or 5.1.2.
5.1.1 Usual procedure.
Weigh, to the nearest mg, 1 g or more of the sample. Place the test sample in a Kjeldahl flask, add 20 ml of sulphuric acid (3.5), shake to impregnate the substance completely with acid and to prevent it from sticking to the sides of the flask, heat and keep at boiling point for 10 minutes. Leave to cool slightly, add 2 ml of nitric acid (3.4), heat gently, leave to cool slightly, add a little more nitric acid (3.4) and bring back to boiling point. Repeat this procedure until a colourless solution is obtained. Cool, add a little water, decant the liquid into a 500 ml graduated flask, rinsing the Kjeldahl flask with hot water. Leave to cool, make up to volume with water, mix and filter.
5.1.2 Samples containing organic substances and free from calcium and magnesium dihydrogen phosphates.
Weigh, to the nearest mg, about 2.5 g of the sample in an ashing crucible (4.1). Mix the test sample until completely merged with 1 g of calcium carbonate (3.1). Ash in the oven at 550C±5C until white or grey ash is obtained (a little charcoal does not matter). Transfer the ash into a beaker. Add 20 ml of water and hydrochloric acid (3.2) until effervescence ceases. Add a further 10 ml of hydrochloric acid (3.2). Place the beaker on a sand bath and evaporate to dryness to make the silica insoluble. Redissolve the residue in 10 ml of nitric acid (3.3) and boil on the sand bath for 5 minutes. Decant the liquid into a 500 ml graduated flask, rinsing the beaker several times with hot water. Leave to cool, make up to volume with water, mix and filter.
5.2 Development of colouration and measurement of optical density.
Dilute an aliquot part of the filtrate obtained by 5.1.1 or 5.1.2 to obtain a phosphorus concentration of not more than 40 vg/ml. Place 10 ml of this solution in a test tube (4.6) and add 10 ml of molybdovanadate reagent (3.6). Mix and leave to stand for at least 10 minutes at 20C. Measure the optical density in a spectrophotometer (4.5), at 430 nm against a solution obtained by adding 10 ml of the molybdovanadate reagent (3.6) to 10 ml of water.
5.3 Calibration curve.
From the standard solution (3.7) prepare solutions containing respectively 5, 10, 20, 30 and 40 vg of phosphorus per ml. Take 10 ml of each of these solutions and add 10 ml of molybdovanadate reagent (3.6). Mix and leave to stand for at least 10 minutes at 20C. Measure the optical density as indicated in 5.2.
Trace the calibration curve by plotting the optical densities against the corresponding quantities of phosphorus. For concentrations between 0-40 vg/ml, the curve will be linear.
6.
Calculation of results
Determine the amount of phosphorous in the test sample by using the calibration curve. Express the result as a percentage of the sample.
7.
Repeatability
The difference between results of two parallel determinations carried out on the same sample should not exceed
3% relative to the higher result, for phosphorus contents of less than 5%; 0.15% in absolute value, for phosphorus contents equal to or greater than 5%.
6.8 DETERMINATION OF POTASSIUM.
1.
Purpose and scope
To determine the content of potassium in feeding stuffs.
2.
Principle
The sample is ashed and the ash dissolved in hydrochloric acid. The potassium content of the solution is determined by flame photometry in the presence of caesium chloride and aluminium nitrate. The addition of these substances largely eliminates interference from disturbing elements.
3.
Reagents
3.1 Hydrochloric acid d : 1.12.
3.2 Caesium chloride.
3.3 Aluminium nitrate, Al(NO3)3.9H2O.
3.4 Potassium chloride, anhydrous.
3.5 Loading agent: dissolve in water 50 g of caesium chloride (3.2) and 250 g of aluminium nitrate (3.3), make up to 1 litre with water and mix. Store in plastic bottles.
3.6 Standard solution of potassium: dissolve in water 1.907 g of potassium chloride (3.4), add 5 ml of hydrochloric acid (3.1), make up to 1 litre with water and mix. Store in plastic bottles. 1 ml of this solution contains 1.00 mg of potassium.
4.
Apparatus
4.1 Platinum, silica or porcelain crucibles for ashing, provided if necessary with lids.
4.2 Electric muffle-furnace with thermostat.
4.3 Flame photometer.
5.
Procedure
5.1 Analysis of sample.
As a general rule, weigh, to the nearest 10 mg approximately 10 g of the sample, place in a crucible (4.1) and ash at 450C for 3 hours. After cooling, transfer the ash quantitatively to a 500 ml graduated flask using 250-300 ml of water and then 50 ml of hydrochloric acid (3.1). When all release of carbon dioxide has ceased, heat the solution and keep at a temperature of about 90C for 2 hours, stirring occasionally. After cooling to room temperature, make up to the mark with water, shake and filter. Transfer to a 100 ml graduated flask an aliquot part of the filtrate containing a maximum of 1.0 mg of potassium, add 10.0 ml of loading agent (3.5), make up to the mark with water and mix. In the case of higher levels of potassium dilute the solution to be analysed in suitable proportions before adding the loading agent.
The table below is given as a guide for a sample of about 10 g.
Assumed potassium content of the sample (%K)
Dilution factor
Aliquot part in ml of
the solution
Up to 0.1
—
50
0.1 to 0.5
—
10
0.5 to 1.0
—
5
1.0 to 5.0
1 : 10
10
5.0 to 10.0
1 : 10
5
10.0 to 20.0
1 : 20
5
Measure by flame photometry at a wavelength of 768 nm. Calculate the results by means of a calibration curve.
5.2 Calibration curve.
Place exactly 10 ml of the standard solution (3.6) in a 250 ml graduated flask, make up to the mark with water and mix. Place in 100 ml graduated flasks exactly 5, 10, 15, 20 and 25 ml of this solution, corresponding respectively to amounts of potassium of 0.2, 0.4, 0.6, 0.8 and 1.0 mg. Complete the series with a blank flask containing no standard solution. Add 10 ml of loading agent (3.5) to each flask, make up to the mark with water and mix. Carry out the measurements as indicated in 5.1. The calibration curve is generally linear up to potassium concentration of 1 mg in 100 ml of solution.
6.
Calculation of results.
Express the result as a percentage of the sample.
7.
Observation
It is not always necessary to add loading agent (3.5) in order to eliminate the interference of disturbing elements.
6.9 DETERMINATION OF SODIUM.
1.
Purpose and scope
To determine the content of sodium in feeding stuffs.
2.
Principle
The sample is ashed and the ash dissolved in hydrochloric acid. The sodium content of the solution is determined by flame photometry in the presence of caesium chloride and aluminium nitrate. The addition of these substances largely eliminates interference from disturbing elements.
3.
Reagents
3.1 Hydrochloric acid, d : 1.12.
3.2 Caesium chloride.
3.3 Aluminium nitrate, Al(NO
3
)
3
.9H
2
O.
3.4 Sodium chloride, anhydrous.
3.5 Loading agent: dissolve in water 50 g of caesium chloride (3.2) and 250 g of aluminium nitrate (3.3), make up to 1 litre with water and mix. Store in plastic bottles.
3.6 Standard solution of sodium: dissolve in water 2.542 g of sodium chloride (3.4) add 5 ml of hydrochloric acid (3.1), make up to 1 litre with water and mix. Store in plastic bottles. 1 ml of this solution contains 1.00 mg of sodium.
4.
Apparatus
4.1 Platinum, silica or porcelain crucibles for ashing, provided if necessary with lids.
4.2 Electric muffle-furnace with thermostat.
4.3 Flame photometer.
5.
Procedure
5.1 Analysis of sample
As a general rule, weigh, to the nearest 10 mg, approximately 10 g of the sample, place in a crucible (4.1) and ash at 450°C for 3 hours. Avoid overheating (ignition). After cooling, transfer the ash quantitatively to a 500 ml graduated flask, using 250-300 ml of water and then 50 ml of hydrochloric acid (3.1). When all release of carbon dioxide has ceased, heat the solution and keep at a temperature of about 90°C for 2 hours, stirring occasionally. After cooling to room temperature, make up to the mark with water, shake and filter. Transfer to a 100 ml graduated flask an aliquot part of the filtrate containing a maximum of 1.0 mg of sodium, add 10.0 ml of loading agent (3.5), make up to the mark with water and mix. In the case of higher levels of sodium, dilute the solution to be analysed in suitable proportions before adding the loading agent. Measure by flame photometry at a wavelength of 589 nm. Calculate the result by means of a calibration curve.
The table below is given as a guide for a sample of about 10 g.
Assumed sodium content of the sample (%Na)
Dilution factor
Aliquot part in ml of
the solution
Up to 0.1
—
50
0.1 to 0.5
—
10
0.5 to 1.0
—
5
1.0 to 5.0
1 : 10
10
5.0 to 10.0
1 : 10
5
10.0 to 20.0
1 : 20
5
5.2 Calibration curve.
Place exactly 10 ml of the standard solution (3.6) in a 250 ml graduated flask, make up to the mark with water and mix. Place in 100 ml graduated flasks exactly 5, 10, 15, 20 and 25 ml of this solution, corresponding respectively to amounts of sodium of 0.2, 0.4, 0.6, 0.8 and 1.0 mg. Complete the series with a blank flask containing no standard solution. Add 10 ml of loading agent (3.5) to each flask, make up to the mark with water and mix. Carry out the measurements as indicated in 5.1. The calibration curve is generally linear up to a sodium concentration of 1 mg in 100 ml of solution.
6.
Calculation of results
Express the result as a percentage of the sample.
7.
Observations
7.1 For products containing more than 4% of sodium, it is preferable to ash the substance for 2 hours in a crucible with a lid. After cooling, add water, bring the ash into suspension by means of a platinum wire, dry and ash again for 2 hours in the crucible with a lid.
7.2 If the sample consists solely of mineral substances, dissolve without prior ashing.
7. MISCELLANEOUS.
7.1 DETERMINATION OF HECTOLITRE WEIGHT.
1.
Purpose and Scope
To determine the hectolitre weight of cereals.
2.
Principle
The bulk density of cereals expressed as kilogram per hectolitre, is calculated from the weight of one bushel (measured in a specified manner) of cereal. Bushel weight is converted to hectolitre weight by multiplying by the factor 1.25.
3.
Apparatus
3.1 Balance and weights conforming to the weights and measures Act 1904.
3.2 Measuring cylinder made of brass or copper with a perforated base an observation window fitted at its lower end and a slot at the upper end for the insertion and withdrawal of the levelling plate (3.3).
3.3 Levelling plate.
3.4 Auxiliary cylinder made of brass or copper.
3.5 A filling cylinder with a double funnel arrangement at its outlet.
3.6 Cylindrical drop weight.
3.7 Disk.
4.
Procedure
Check that the weights pan exactly counterbalances the measuring cylinder (3.2) and the drop weight (3.6). Place the measuring cylinder (3.2) on the disk (3.7). Insert the levelling plate (3.3) into the slot at the top of the measuring cylinder (3.2) and place the drop weight (3.6) on top of it. Place auxiliary cylinder (3.4) on top of (3.2).
Fill the filling cylinder (3.5) with cereal to the level of the outlet hole (do not tap the tube). Place on top of cylinder (3.3) allow the contents to empty completely into (3.4) and then remove cylinder (3.5). Withdraw the levelling plate (3.3) completely and reinsert after the drop weight (3.6) has fallen to the bottom of the measuring cylinder (3.2). Remove the auxiliary cylinder (3.4) and discard the surplus cereal remaining above the levelling plate (3.3).
Remove the levelling plate (3.3) and weigh the measuring cylinder (3.2) together with the drop weigh (3.6) and the cereal, to obtain the bushel weight.
5.
Calculation of results
To convert bushel weight to hectolitre weight, multiply the weight obtained in the preceding paragraph by 1.25.
6.
Repeatability
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% in absolute value.
7.2 DETERMINATION OF TOTAL SOLIDS.
1.
Purpose and Scope
To determine the total solids content of milk products.
2.
Principle
A known quantity of sample is dried at a constant temperature and to a constant weight. The residue after drying constitutes the total solids.
3.
Apparatus
3.1 Non-corrodible, flat metal dishes about 2 cm deep and approximately 6–8 cm in diameter with well fitting lids.
3.2 Air drying oven at 102°C±2°C.
3.3 Water bath.
4.
Procedure
Place dish (3.1) in a drying oven (3.2) for approximately 30 minutes. Cool in a dessicator and weigh accurately. Weigh to the nearest mg approximately 3 g of sample directly into the dish and place the uncovered dish on a boiling water bath for 30 minutes. Transfer to an oven (3.2) and leave for 2 hours. Remove to a desiccator, cool and weigh. Dry in the oven (3.2) for a further hour, cool and weigh. Repeat the drying and weighing until the difference between two successive weighings is not more than 0.5 mg.
5.
Calculation of results
Express the result as a percentage of the sample.
7.3 DETERMINATION OF THE UREASE ACTIVITY OF PRODUCTS DERIVED FROM SOYA BEANS.
1.
Purpose and scope
To estimate the urease activity of products derived from soya beans and to show whether these products have been cooked for a sufficient length of time.
2.
Principle
Urease activity is estimated by the amount of ammoniacal nitrogen liberated per 1 g of product per minute at 30°C from a solution of urea.
3.
Reagents
3.1 Hydrochloric acid, 0.1 N.
3.2 Sodium hydroxide solution, 0.1 N.
3.3 Loading agent of phosphate 0.05 M, containing, per 1000 ml, 4.45 g of disodium phosphate (Na
2
HPO
4
.2H
2
O) and 3.40 g of monopotassium phosphate (KH
2
PO
4
).
3.4 Freshly-prepared urea loading agent containing 30.0 g of urea per 1000 ml of loading agent (3.3); pH 6.9-7.0.
4.
Apparatus
4.1 Potentiometric titration apparatus or high sensitivity pH-meter (0.02 pH) with magnetic stirrer.
4.2 Water bath fitted with thermostat set at 30°C exactly.
4.3 Test tubes with ground glass stoppers.
5.
Procedure
Crush about 10g of the sample (e.g. in a coffee mill) so that it passes through a sieve with a mesh of 0.2 mm. Weigh 0.2 g of the crushed sample to the nearest mg, place in a test tube with a ground glass stopper and add 10 ml of urea loading agent (3.4). Stopper immediately and shake vigorously. Place the tube in a water bath (4.2), set at 30°C exactly and keep there for exactly 30 minutes. Immediately add 10 ml of 0.1 N hydrochloric acid (3.1) cool rapidly to 20°C and transfer the contents of the tube quantitatively to a titration vessel, rinsing twice with 5 ml of water. Using a glass electrode (4.1) titrate immediately and rapidly to pH 4.7 with the 0.1 N sodium hydroxide solution (3.2) by electrometry.
Carry out a blank test as follows
Quickly place a sample of 0.2 g, weighed to the nearest mg, in a test tube with a ground glass stopper, add 10 ml of 0.1 N hydrochloric acid (3.1) and then 10 ml of urea loading agent (3.4). Cool the tube immediately in ice-water and leave there for 30 minutes. Under the conditions indicated above, transfer the contents of the tube to the titration vessel using the 0.1 N sodium hydroxide solution (3.2) up to pH 4.7.
6.
Calculation of results
The urease activity is calculated by using the formula
where
a=ml of 0.1 N sodium hydroxide solution consumed by the sample;
b=ml of 0.1 N sodium hydroxide solution consumed in the blank test;
E=weight of the sample in grams.
7.
Observations
7.1 This method is suitable for a urease activity of up to 1 mg of N/g/minute at 30°C. For more active products, the size of the sample can be reduced to 50 mg.
7.2 Products containing more than 10% of crude fatty substance must first be defatted cold.
7.4 DETERMINATION OF VITAMIN A (RETINOL)
1.
Purpose and scope
To determine the content of Vitamin A in feeding stuffs, concentrates and premixtures. The lower limit of the determination is 10,000 IU/kg for highly pigmented feeds and 4.000 IU/kg for others
1
. Products are classified in two groups, according to their presumed Vitamin A contents
1
1 IU = 0.3μg of Vitamin A.
Group A: contents lower than 200,000 IU/kg;
Group B: contents equal to or greater than 200,000 IU/kg.
2.
Principle
The sample is hydrolised in hot ethanolic potassium hydroxide solution and in the presence of an antioxidant or in a nitrogen atmosphere. The mixture is extracted with 1, 2-dichlorethane. The extract is evaporated to dryness and treated with light petroleum. The solution is chromatographed on a column of aluminium oxide (for Group B products, chromatography is only required in certain cases). For Group A products the Vitamin A is determined by spectrophotometry at 610 nm after development of a coloured complex according to the Carr-Price reaction; for Group B products by spectrophotometry in the UV at 325 nm.
3.
Reagents
(a) used for analysing products of Groups A and B.
3.1 Ethanol, 96%(v/v).
3.2 Sodium ascorbate solution, 10% (w/v) or
3.3 Purified nitrogen.
3.4 Potassium hydroxide solution, 50% (w/v).
3.5 Potassium hydroxide solution, 1 N.
3.6 Potassium hydroxide solution. 0.5 N.
3.7 1, 2-dichlorethane.
3.8 Light petroleum, boiling range: 40-60°C; if necessary, purify as follows: stir 1,000 ml light petroleum with 20 ml lots of concentrated sulphuric acid until the acid remains colourless. Remove the acid and wash the light petroleum successively with 500 ml water, twice with 250 ml of 10% (w/v) sodium hydroxide solution and three times with 500 ml water. Remove the aqueous layer, dry the light petroleum for 1 hour over active carbon and anhydrous sodium sulphate, filter and distil.
3.9 Aluminium oxide, standardised according to Brockmann: ash for 8 hours at 750°C. cool in a desiccator and keep in a brown glass bottle fitted with a ground glass stopper. Before use in chromatography moisten as follows: place in a brown glass bottle 10 g aluminium oxide and 0.9 ml water, seal with a stopper, reheat for 5 minutes in a boiling water bath while shaking. Leave to cool. Verify the activity of the aluminium thus prepared by subjecting a known quantity of Vitamin A (3.17) (approximately 500 IU) to the procedure of 5.3 and 5.4 and checking recovery.
3.10 Basic aluminium oxide, degree of activity 1 (Woelm, Merck or equivalent).
3.11 Diethyl ether: remove peroxides and traces of water by chromatography on a column of basic aluminium oxide (3.10). (25 g aluminium oxide per 250 ml diethyl ether).
3.12 Light petroleum solutions (3.8) containing 4, 8, 12, 16 and 20% (v/v) diethyl ether (3.11).
3.13 Sodium sulphide solution 0.5 molar in 70% (v/v) glycerine, prepared from sodium sulphide.
(b) used exclusively for analysing Group A products.
3.14 Crystallizable benzene.
3.15 Chloroform: remove the ethanol, phosgene and traces of water by chromatography on a column of basic aluminium oxide (3.10) (50 g aluminium oxide per 200 ml chloroform; it is advisable to chromatograph the first 50 ml of the eluate a second time).
3.16 Carr-Price reagent: stir approximately 25 g antimony trichloride (kept in a desiccator) with 100 ml chloroform (3.15) until the solution is saturated. A slight deposit of antimony trichloride causes no problem. Add 2 ml acetic anhydride. Keep in a refrigerator in a brown glass bottle with glass stopper. The solution keeps for several weeks.
3.17 Vitamin A — standardized spectrophotometrically.
(c) used exclusively for analysing Group B products.
3.18 Isopropanol, for chromatography.
8. (1) Subject to paragraphs (2) and (3) of this Regulation, feedingstuffs shall be marketed only if the following particulars, which shall be clearly visible, legible and indelible, are set out on the package or container or on a label attached thereto
(
a
) in case a feeding stuff is a straight feedingstuff—
(i) unless the feedingstuff is marketed to a person described in Regulation 5 (
e
) of these Regulations, the words "straight feedingstuff",
(ii) the particulars specified in column (4) of Part I of the First Schedule to these Regulations opposite where the name of the feedingstuff appears in column (2) thereof together with the words or word, as may be appropriate, used as its name in the said column (2),
(iii) the particulars specified in paragraphs (
e
) and (
f
) of Article 7 (1) of the Directive of 1976,
(iv) where appropriate, the particulars specified in paragraphs 1.1 and 2 of Part A of the Annex to the Directive of 1976 together with, in case the feedingstuff is a feedingstuff listed in item 2.9.2 of Part B of the said Annex, the particulars specified in paragraph 1.3 of the said Part A,
(v) in case the feedingstuff is a feedingstuff described in paragraph (
e
) of Regulation 5 of these Regulations, the declaration required by the said Regulation 5.
(
b
) in case the feedingstuff is a compound feedingstuff—
(i) where the feedingstuff is a complete feedingstuff, the description "complete feedingstuff",
(ii) where the feedingstuff is a complementary feedingstuff—
(I) if it is a complementary mineral feedingstuff, the description "complementary mineral feedingstuff"
(II) if it is a complementary molassed feedingstuff, the description "complementary molassed feedingstuff"
and
(III) if it is neither a complementary mineral feedingstuff nor a complementary molassed feedingstuff, the description "complementary feedingstuff",
(iii) the particulars listed in paragraphs (
b
), (
c
), (
d
), (
f
) and (
g
) of Article 5 (1) of the Directive of 1979,
(iv) except in the case of feedingstuffs which are intended for pet animals, the date of manufacture,
(v) in case the feedingstuff is a milk replacer (suckling feed), the milk powder content,
(vi) the declarations listed in column (2) of part III of the First Schedule to these Regulations opposite the mention in column (1) of that Part of the relevant type of feedingstuff, and
(vii) the declaration required by Regulation 6 (1) of these Regulations and referred to in Regulation 6 (2) (
e
) (iv) of these Regulations
Provided that
(I) in case of small quantities of compound feedingstuffs intended for the final user, in lieu of complying with the requirements of subparagraph (
b
) of this paragraph, the particulars specified in that subparagraph may be brought to the purchasers' attention by means of an appropriate notice; and
(II) in case the feedingstuff is constituted from no more than three ingredients, the particulars referred to in subparagraph (
b
) and, where appropriate, either or both of the following, namely, subparagraphs (
c
) or (
d
) of the said Article 5 (1) shall not be required if the ingredients used in the feedingstuff appear clearly in a description of the feedingstuff contained in the aforesaid particulars.
(2) (
a
) In relation to straight feedingstuffs marketed in bulk, paragraph (1) of this Regulation shall be construed as requiring the particulars specified in that paragraph to be set out in an accompanying document.
(
b
) In relation to compound feedingstuffs duly marketed in tankers or similar vehicles or otherwise in bulk, paragraph (1) of this Regulation shall be construed as requiring the particulars to be set out in an accompanying document.
(3) The declarations referred to in paragraph (1) (
b
) of this Regulation shall each comply with the requirements of point 1 of the Annex to the Directive of 1979.
9. (1) A person who markets a straight feedingstuff shall not put on the package, container or label thereof or on an accompanying document, in conjunction with any of the particulars set out in compliance with the requirements of Regulation 8 (1) of these Regulations, additional particulars other than the following, namely—
(
a
) the particulars listed in Article 7 (5) of the Directive of 1976 (other than paragraph (
g
)),
and
(
b
) the declarations listed in column (5) of Part I of the First Schedule to these Regulations opposite the mention in column (2) thereof of the relevant feedingstuff.
(2) A person who markets a compound feedingstuff shall not put on the package, container, label or accompanying document thereof in conjunction with the particulars set out in compliance with the requirements of Regulation 8 (1) (
b
) of these Regulations additional particulars other than—
(
a
) the particulars listed in Article 5 (5) of the Directive of 1979,
(
b
) the trade name of the product,
(
c
) each of the ingredients in descending order of its proportion by weight,
(
d
) the declarations listed in column (3) of Part III of the First Schedule to these Regulations opposite the mention in column (1) thereof of the relevant type of feedingstuff,
(
e
) where it is appropriate, an indication that the compound feedingstuff conforms to a standard or standards regarding analytical characteristics which for the time being stands or stand recommended by the Minister in relation to a particular category or particular categories of compound feedingstuffs.
(3) Paragraph (3) of Regulation 8 of these Regulations shall apply to a declaration, other than a declaration referred to in paragraph (1) (
b
) of that Regulation, which pursuant to these Regulations is put on a package, container, label or accompanying document as it applies to a declaration so referred to.
10. (
a
) Where information referred to in Article 7 (7) of the Directive of 1976 appears on any packaging, container, label or accompanying document to which the said Article 7 (7) relates, the information shall be given in accordance with the requirements of that Article.
(
b
) Where information referred to in Article 5 (8) of the Directive of 1979 appears on any packaging, container, label or accompanying document to which the said Article 5 (8) relates, the information shall be shown in accordance with the requirements of that Article.
11. (1) Where a person has on his premises any feedingstuff (whether straight or compound) which he has purchased and which he proposes to use in the course of his farming operations, he may apply to the Minister to have a sample thereof taken for analysis.
(2) An application under this Regulation shall be—
(
a
) made within the period of thirty days beginning on the date on which the feedingstuff to which the application relates was delivered to the applicant, and
(
b
) accompanied by a fee which shall be calculated by reference to the rates specified in the Second Schedule to these Regulations.
(3) Where an application is made under this Regulation, an authorised person shall, subject to paragraph (4) of this Regulation—
(
a
) take and deal with a sample of the relevant feedingstuff according to the methods described in the Annex to Commission Directive No. 76/371/EEC of 1 March, 1976
1
,
1
O.J. No. L102/1, 15 April 1976.
and
(
b
) give or cause to be given, or sent by registered post or by such other method as for the time being stands approved of for the purposes of this paragraph by the Minister, to the State Chemist and to the person whose name or trade name has been given pursuant to paragraph (
f
) of Article 7 (1) of the Directive of 1976, or, as may be appropriate, paragraph (
f
) of Article 5 (1) of the Directive of 1979, one of the final samples prepared pursuant to the requirements of subparagraph (
a
) of this paragraph.
(4) Where an application is made under this Regulation, an authorised person may, if he thinks fit, decline to take a sample if—
(
a
) he is not satisfied that the applicant has purchased the feedingstuff to which the application relates,
or
(
b
) he is not satisfied that the applicant proposes to use such feedingstuff in the course of his farming operations,
or
(
c
) he is not satisfied that such feedingstuff as presented for sampling is fairly representative of the feedingstuff as delivered to the applicant,
or
(
d
) the applicant does not furnish such information relating to such feedingstuff as the authorised person may reasonably require.
(5) Where the State Chemist receives a sample taken in pursuance of an application under this Regulation, he shall in making an analysis thereof comply with any of the requirements set out in Part I of the Third Schedule to these Regulations, as apply in the particular case and send to the applicant and to the person (other than the State Chemist) referred to in paragraph (3) (
b
) of this Regulation a certificate, in the form set out in Part II of the said Third Schedule, of the result of the analysis.
(6) Subject to paragraph (8) of this Regulation, all fees under this Regulation shall be paid into or disposed of for the benefit of the Exchequer in accordance with the directions of the Minister for Finance and, accordingly, the Public Offices Fees Act, 1879, shall not apply in respect thereof.
(7) Nothing in this Regulation shall be construed as requiring the State Chemist to make a test, examination or analysis regarding the presence in or absence from a sample given or sent to him pursuant to these Regulations of any particular substance, product or other thing, if in his opinion there is not in relation to such presence or absence a method of testing, examination or analysis which is sufficiently reliable or if there is not available to the State Chemist the apparatus or other means by which such a test, examination or analysis could be made.
(8) In any case in which he considers it proper so to do (not being a case in which the applicant has received a certificate under this Regulation), the Minister may refund a fee paid in relation to an application under this Regulation.
(9) For the purpose of this Regulation a feedingstuff shall not be regarded as having been delivered to a purchaser until it arrives at the destination to which it is consigned whether the consignment is by direction of the supplier or the purchaser.
12. Every person who carries on, or is employed in connection with, the business of manufacturing, importing or marketing feedingstuffs shall—
(
a
) keep records of his transactions in such feedingstuffs;
(
b
) produce at the request of an authorised person any books, documents or records relating to such business which are in the possession or under the control of such person;
(
c
) permit an authorised person to inspect and take extracts from such books, documents or records and give to the person any information which he may reasonably require in relation to any entries therein;
(
d
) afford to an authorised person reasonable facilities for inspecting the stock of any feedingstuff which is for the time being on any premises on which such person carries on such a business;
(
e
) give to an authorised person any information he may reasonably require in relation to such transactions including in particular information which he may reasonably require regarding any feedingstuff which is specified by him.
13. Where an authorised person is satisfied that a feedingstuff which is placed on the market or which he believes will be placed on the market, does not comply with any one or more of the requirements of these Regulations, he may require either or both of the following persons, namely, the person who appears to him to have for the time being possession or control of the feedingstuff and the person whose name or trade name has been given pursuant to paragraph (
f
) of Article 7 (1) of the Directive of 1976, or, as may be appropriate, paragraph (
f
) of Article 5 (1) of the Directive of 1979, to take such steps as are necessary to ensure that it does not continue to be placed on the market, or, as may be appropriate, is not placed on the market until such authorised person is satisfied that the requirement is complied with.
14. In addition to the powers conferred by Regulation 12 of these Regulations an authorised person may at all reasonable times enter an inspect any premises or any railway wagon, vehicle, ship, vessel or aircraft, or other thing used to transport (e.g. a container), in which he has reasonable grounds for believing that any feedingstuff is being marketed, manufactured for sale, imported, stored or kept for sale, sold or transported and may examine and take samples of any feedingstuff which he finds in the course of his inspection.
15. (1) Where in any proceedings for an offence in which a contravention of these Regulation is alleged the defendant claims that the feedingstuff to which the alleged offence relates was exempted from these Regulations, by reason of paragraph (
a
) or (
b
) of Regulation 3 of these Regulations, the onus of proving that such feedingstuff was so exempted shall be on the defendant.
(2) In any proceedings for an offence under these Regulations, evidence of the result of any test, examination or analysis of, or of any report on, a sample taken under these Regulations may be given if, and only if, it is proved that—
(
a
) the sample was taken and dealt with in accordance with the methods described in the Annex to Commission Directive No. 76/371/EEC of 1 March, 1976
1
,
1
O.J. No. L102/1, 15 April 1976.
(
b
) before the proceedings were instituted one of the final samples prepared pursuant to the requirements of the said Annex was given or caused to be given to the defendant or sent or given to him by registered post or by such other method as stands approved of for the purposes of paragraph (
b
) of Regulation 11 (3) of these Regulations by the Minister, and
(
c
) the test, examination or analysis was carried out in accordance with such of the requirements (if any) specified in Part I of the Third Schedule to these Regulations as applied in the particular case.
(3) In any legal proceedings, other than proceedings to which paragraph (2) of this Regulation applies, evidence, being evidence which relates to an issue regarding the accuracy of a declaration required to be made by these Regulations, of the result of any test, examination or analysis of a sample of a feedingstuff may be given if, and only if, it is proved that—
(
a
) the sample was taken and dealt with in accordance with the methods referred to in paragraph (2) (
a
) of this Regulation,
(
b
) before the proceedings were instituted one of the final samples prepared pursuant to the requirements of the Annex to Commission Directive No. 76/371/EEC of 1 March, 1976, was given or caused to be given to the defendant, or sent or given to him by registered post or by any other method which is a method referred to in paragraph (2) (
b
) of this Regulation, and
(
c
) the test, examination or analysis was carried out in accordance with such of the requirements referred to in paragraph (2) (
c
) of this Regulation as applied in the particular case.
16. The Minister shall furnish an authorised person with a certificate of his appointment and, when exercising any powers conferred by these Regulations, the authorised person shall if requested by any person affected, produce the certificate to that person.
17. In any legal proceedings the production of a certificate in the form specified in Part II of the Third Schedule to these Regulations and purporting to be signed by the State Chemist shall, without proof of any signature on the certificate or that the signatory was the proper person to sign it, be sufficient evidence of the facts stated in the certificate and of the analysis to which it relates having been carried out in accordance with such of the requirements (if any) specified in Part I of the said Third Schedule as applied in the particular case.
18. (1) Any person who—
(
a
) contravenes Regulation 5 or 6 of these Regulations,
(
b
) in marketing a feedingstuff—
(i) contravenes Regulation 9 of these Regulations,
(ii) fails to comply with a requirement of Regulation 8 or 10 of these Regulations, or of the said Regulation 8, as applied by Regulation 9 (3) of these Regulations, other than a requirement as regards the disclosed composition of a feedingstuff,
(
c
) contravenes any of the provisions of Regulation 12 of these Regulations,
(
d
) fails to comply with a requirement made of him under Regulation 13 of these Regulations,
(
e
) obstructs or interferes with an authorised person in the course of exercising a power conferred on him under Regulation 14 of these Regulations,
shall be guilty of an offence.
(2) Any person who in marketing a feedingstuff fails to comply with a requirement of Regulation 8 or 9 of these Regulations as regards the disclosed composition of a feedingstuff shall, subject to the limits of error specified in the Fourth Schedule to these Regulations, be guilty of an offence.
(3) If any person fraudulently—
(
a
) tampers with any thing so as to procure that any sample taken pursuant to Regulation 11 or 14 of these Regulations does not correctly represent the substance sampled, or
(
b
) tampers or interferes with any sample taken under these Regulations,
he shall be guilty of an offence.
(4) Any person guilty of an offence under these Regulations shall be liable on summary conviction to a fine not exceeding £500, or, at the discretion of the court, to imprisonment for a term not exceeding six months.
(5) An offence under these Regulations may be prosecuted by the Minister.
FIRST SCHEDULE
PART I
STRAIGHT FEEDINGSTUFFS
Name of Feedingstuff
Description
Compulsory
Declarations
Optional
Declarations
Compositional
Requirements
1
2
3
4
5
6
1.
Oil cakes and meals
1.1
Macoya palm kernel expeller.
By-product of oil manufacture, obtained by pressing from seeds separated from their pulp of the following species of Macoya palm: Acrocomia sclerocarpa Mart, and Acrocomia totai Mart.
Crude protein
Crude fibre
Crude oil and fat
Crude ash
Moisture>12%
Moisture
Ash insoluble in HCl Max 2%
1.2
Macoya extracted palm kernal.
By-product of oil manufacture, obtained by extraction from seeds of Macoya palm separated from their pulp.
Crude protein
Crude fibre
Crude ash
Moisture>12.5%
Crude oil and fat Moisture
Ash insoluble in HCl Max. 2%
1.3
Macoya palm pulp.
By-product of oil manufacture, obtained by pressing from pulp of Macoya palm.
Crude protein
Crude fibre
Crude oil and fat
Crude ash
Moisture>12%
Moisture
Ash insoluble in HCl Max. 2%
1.4
Decorticated ground nut expeller.
By-product of oil manufacture, obtained by pressing from decorticated ground nuts(species Arachis hypogaea and other species of Arachis).
Crude protein
Crude fibre
Crude oil and fat
Crude ash
Moisture>12%
Moisture
Ash insoluble in HCl Max. 2%
1.5
Extracted decorticated groundnut.
By-product of oil manufacture, obtained by extraction from decorticated groundnut seeds.
Crude protein
Crude fibre
Crude ash
Moisture>12.5%
Crude oil and fat Moisture
Ash insoluble in HCl Max. 2%
1.6
Partly-decorticated groundnut expeller.
By-product of oil manufacture, obtained by pressing from partly-decorticated ground nut seeds.
Crude protein
Crude fibre
Crude oil and fat
Crude ash
Moisture>12%
Moisture
Ash insoluble in HCl Max. 2%
1.7
Extracted partly-decorticated groundnut.
By-product of oil manufacture, obtained by extraction from partly-decorticated ground nut seeds.
Crude protein
Crude fibre
Crude ash
Moisture> 12.5%
Crude oil and fat Moisture
Ash insoluble in HCl Max. 2%
1.8
Rape seed expeller.
By-product of oil manufacture, obtained by pressing from seeds of colza Brassica napus L.ssp. oleifera (Metzg) Sinsk., of Indian sarson Brassica napus L. var. glauca (Roxb.) O.E. Schulz and of rape Brassica campestris L.ssp. oleifera (Metzg.) Sinsk.
Crude protein
Crude fibre
Crude oil and fat
Crude ash
Moisture> 12%
Moisture
Ash insoluble in HCl Max. 2%
1.9
Extracted rape seed.
By-product of oil manufacture, obtained by extracting from seeds of colza, Indian sarson or rape.
Crude protein
Crude fibre
Crude ash
Moisture> 12.5%
Crude oil and fat Moisture
Ash insoluble in HCl Max. 2%
1.10
Copra expeller.
By-product of oil manufacture, obtained by pressing from copra, the dried kernel (endosperm) and testa of the coconut palm, Cocos nucifera L.
Crude protein
Crude fibre
Crude oil and fat
Crude ash
Moisture> 12%
Crude oil and fat Moisture
Ash insoluble in HCl Max. 2%
1.11
Extracted copra.
By-product of oil manufacture, obtained by extraction from copra, the dried kernel (endosperm) and testa of the coconut palm.
Crude ash
Crude protein
Crude fibre
Moisture> 12.5%
Crude oil and fat Moisture
Ash insoluble in HCl Max. 2%
1.12
Palm kernel expeller.
By-product of oil manufacture, obtained by pressing from palm nuts, from which as much as possible of the hard shell has been removed, of the following species of oil palm: Elaeis guineensis Jacq., Corozo oleifera (H.B.K.) L. H. Bailey (Elaeis melanococca-auct.)
Crude protein
Crude fibre
Crude oil and fat
Crude ash
Moisture> 12%
Moisture
Ash insoluble in HCl Max. 2%
1.13
Extracted palm kernel.
By-product of oil manufacture, obtained by extraction from palm nuts of the species of oil palm from which as much as possible of the hard shell has been removed.
Crude protein
Crude fibre
Crude ash
Moisture> 12.5%
Crude oil and fat
Moisture
Ash insoluble in HCl Max. 2%
1.14
Soya expeller.
By-product of oil manufacture, obtained by pressing from soya beans (the seed of the species Glycine max. (L.) Merr.).
Crude protein
Crude fibre
Crude oil and fat
Crude ash
Moisture> 12%
Moisture
Ash insoluble in HCl Max. 2%
1.15
Extracted toasted soya.
By-product of oil manufacture, obtained from soya bean seeds by extraction and appropriate heat treatment.
Crude ash
Crude protein
Crude fibre
Moisture> 12.5%
Crude oil and fat
Moisture
Ash insoluble in HCl Max. 2%
1.16
Extracted toasted hulled soya seeds.
By-product of oil manufacture, obtained from hulled soya bean seeds by extraction and appropriate heat treatment.
Crude protein
Crude fibre
Crude ash
Moisture> 12.5%
Crude oil and fat
Moisture
Ash insoluble in HCl Max. 2%
1.17
Decorticated cotton seed expeller.
By-product of oil manufacture, obtained by pressing from seeds of cotton belonging to the genus Gossypium spp. from which the fibres and husks have been removed.
Crude protein
Crude fibre
Crude oil and fat
Crude ash
Moisture> 12%
Moisture
Ash insoluble in HCl Max. 2%
1.18
Extracted decorticated cotton seed.
By-product of oil manufacture, obtained by extraction from seeds of cotton from which the fibres have been removed.
Crude protein
Crude fibre
Crude ash
Moisture> 12.5%
Crude oil and fat
Moisture
Ash insoluble in HCl Max. 2%
1.19
Partly-decorticated cotton seed expeller.
By-product of oil manufacture, obtained from seeds of cotton from which the fibres and part of the husks have been removed.
Crude protein
Crude fibre
Crude oil and fat
Crude ash
Moisture> 12%
Moisture
Ash insoluble in HCl Max. 2%
1.20
Extracted, partly-decorticated cotton seed.
By-product of oil manufacture, obtained by extraction from seeds of cotton from which the fibres and part of the husks have been removed.
Crude protein
Crude fibre
Crude ash
Moisture>12.5%
Crude oil and fat
Moisture
Ash insoluble in HCl Max. 2%
1.21
Expeller or extracted niger seed.
By-product of oil manufacture, obtained by presenting seeds of the niger plant Guizotia abyssinica (L.f.) Cass.
Crude protein
Crude fibre
Crude oil and fat
Crude ash
Moisture> 12%
Moisture
Ash insoluble in HCl Max. 3.4%
1.22
Decorticated sunflower seed expeller.
By-product of oil manufacture, obtained by pressing from seeds of the sunflower Helianthus annuus L. from which as much as possible of the husk has been removed.
Crude protein
Crude fibre
Crude oil and fat
Crude ash
Moisture> 12%
Moisture
Ash insoluble in HCl Max. 2%
1.23
Extracted decorticated sunflower seed.
By-product of manufacture, obtained by extraction from seeds of the sunflower from which the husks have been removed as far as possible.
Crude protein
Crude fibre
Crude ash
Moisture> 12.5%
Crude oil and fat
Moisture
Ash insoluble in HCl Max. 2%
1.24
Partly-decorticated sunflower seed expeller.
By-product of oil manufacture, obtained by pressing from seeds of the sunflower from which part of the husks have been removed.
Crude ash
Crude protein
Crude fibre
Crude oil and fat
Moisture> 12%
Moisture
Ash insoluble in HCl Max. 2%
1.25
Extracted Partly-decorticated sunflower seed.
By-product of oil manufacture, obtained by extraction from seeds of the sunflower from which part of the husks have been removed.
Crude protein
Crude fibre
Crude ash
Moisture> 12.5%
Crude oil and fat
Moisture
Ash insoluble in HCl Max. 2%
1.26
Linseed expeller.
By-product of oil manufacture, obtained by pressing from linseed, Linum usitatissimum L.
Crude protein
Crude fibre
Crude oil and fat
Crude ash
Moisture> 12%
Moisture
Ash insoluble in HCl Max. 2%
1.27
Extracted linseed.
By-product of oil manufacture, obtained by extraction from linseed.
Crude protein
Crude fibre
Crude ash
Moisture> 12.5%
Crude oil and fat
Moisture
Ash insoluble in HCl Max. 2%
1.28
Babassu palm nut expeller.
By-product of oil manufacture, obtained by pressing from palm nuts, from which as much as possible of the hard shell has been removed, of the Brazilian Babassu palms Orbignya oleifera Burr and other species of Orbignya.
Crude protein
Crude fibre
Crude oil and fat
Crude ash
Moisture> 12%
Moisture
Ash insoluble in HCl Max. 2%
1.29
Rice germ expeller.
By-product of oil manufacture, obtained by pressing from germ of rice Oryza sativa L. to which parts of the endosperm and tegument still adhere.
Crude protein
Crude fibre
Crude oil and fat
Crude ash
Moisture> 12%
Moisture
Ash insoluble in HCl Max. 2%
1.30
Extracted brown rice germ.
By-product of oil manufacture, obtained by extraction from germ of rice to which parts of the endosperm and tegument still adhere.
Crude protein
Crude fibre
Crude ash
Moisture> 12.5%
Crude oil and fat
Moisture
Ash insoluble in HCl Max. 2%
1.31
Sesame seed expeller.
By-product of oil manufacture, obtained by pressing from seeds of the sesame plant, Sesamum indicum L.
Crude protein
Crude fibre
Crude oil and fat
Crude ash
Moisture> 12%
Moisture
Ash insoluble in HCl Max. 5%
1.32
Extracted sesame seed.
By-product of oil manufacture, obtained by extraction from seeds of the sesame plant.
Crude protein
Crude fibre
Crude ash
Moisture> 12.5%
Crude oil and fat
Moisture
Ash insoluble in HCl Max. 5%
1.33
Extracted cocoa bean.
By-product of oil manufacture, obtained by extraction from dried and roasted cocoa bean seeds Theobroma Cacao L. from which as much as possible of the husk has been removed.
Crude protein
Crude fibre
Crude ash
Moisture> 12.5%
Crude oil and fat
Moisture
Ash insoluble in HCl Max. 2%
1.34
Wheat germ expeller.
By-product of oil manufacture, obtained by pressing from wheat germ of the species Triticum aestivum L., Triticum durum Desf. and from other cultivated species of husked wheat or from screened husked grains of spelt of the species Triticum spelta L., Triticum dicoccum Schrank, Triticum monococcum L., to which parts of the endosperm and tegument still adhere.
Crude protein
Crude fibre
Crude oil and fat Crude ash
Moisture> 12%
Moisture
Ash insoluble in HCl Max. 2%
1.35
Maize germ expeller (by-product of maize milling).
By-product of oil manufacture, obtained by pressing and by a dry process, from maize germ Zea mays L. to which parts of the endosperm and testa still adhere.
Crude protein
Crude fibre
Crude oil and fat Crude ash
Moisture> 12.5%
Starch
Moisture
Ash insoluble in HCl Max. 2%
1.36
Extracted maize germ (by-product of maize milling).
By-product of oil manufacture, obtained by extraction and by a dry process, from maize germ to which parts of the endosperm and testa still adhere.
Crude protein
Crude fibre
Crude ash
Moisture> 12.5%
Crude oil and fat Starch
Moisture
Ash insoluble in HCl Max. 2%
1.37
Maize germ expeller (by-product of the starch industry).
By-product of oil manufacture, obtained by pressing and by a wet process, from maize germ to which parts of the endosperm and testa still adhere.
Crude protein
Crude fibre
Crude oil and fat
Crude ash Moisture> 12.5%
Moisture
Ash insoluble in HCl Max. 2%
1.38
Extracted maize germ (by-product of the starch industry).
By-product of oil manufacture, obtained by extraction and by a wet process, from maize germ to which parts of the endosperm and testa still adhere.
Crude protein
Crude fibre
Crude ash
Moisture> 12.5%
Crude oil and fat
Moisture
Ash insoluble in HCl Max. 2%
1.39
Olive pulp meal.
By-product of oil manufacture, obtained by extraction from fruits of the olive tree Olea Europa L. free as far as possible from fragments of stone.
Crude protein
Crude fibre
Crude ash
Moisture> 12.5%
Crude oil and fat
Moisture
Ash insoluble in HCl Max. 2%
2.
Products and by-products of the processing of vegetable substances
2.1
by-product of milling
2.1.1
Wheat bran.
By-product of flour manufacture, obtained from screened husked grains of wheat or spelt. It consists principally of fragments of the outer skins, and of particles of grain from which the greater part of the endosperm has been removed.
Crude fibre
Crude ash
Moisture> 14%
Moisture
Ash insoluble in HCl Max. 2%
2.1.2
Wheat feed.
By-product of flour manufacture, obtained from screened husked grains of wheat or spelt. It consists principally of fragments of the outer skins and of particles of grain from which less of the endosperm has been removed than in wheat bran.
Crude fibre
Starch
Crude ash
Moisture> 14%
Moisture
Ash insoluble in HCl Max. 2%
2.1.3
Wheat middlings.
By-product of flour manufacture, obtained from screened husked wheat or spelt. It consists principally of particles of endosperm with fine fragments of the outer skins and some grain waste.
Crude fibre
Starch
Crude ash
Moisture> 14%
Moisture
Ash insoluble in HCl Max. 2%
2.1.4
Wheat germ.
By-product of milling consisting essentially of wheat germ, rolled or otherwise, to which fragments of endosperm and outer skin still adhere.
Crude fibre
Crude protein
Crude oil and fat Crude ash
Moisture> 12%
Moisture
Ash insoluble in HCl Max. 2%
2.1.5
Rye bran.
By-product of flour manufacture, obtained from screened rye secale cereale L. It consists principally of fragments of the outer skins, and of particles of grain from which most of the endosperm has been removed.
Crude fibre
Crude ash
Moisture> 14%
Moisture
Ash insoluble in HCl Max. 2%
2.1.6
Rye feed.
By-product of flour manufacture, obtained from screened rye. It consists principally of fragments of the outer skins, and of particles of grain from which less of the endosperm has been removed than in rye bran.
Crude fibre
Starch
Crude ash
Moisture> 14%
Moisture
Ash insoluble in HCl Max. 2%
2.1.7
Rye screenings (rye meal).
By-products of flour manufacture, obtained from screened rye. It consists principally of particles of endosperm, with fine fragments of the outer skins and some grain waste.
Crude fibre
Starch
Crude ash
Moisture> 14%
Moisture
Ash insoluble in HCl Max. 2%
2.2
Products and by-products of the manufacture of flakes, groats and husked grain.
2.2.1
Husked oat sharps (middlings).
By-product, rich in starch, obtained during the processing of screened, husked, oats Avena sativa L. and other cultivated species of oats into oat groats or sifted oatmeal.
Crude fibre
Starch
Crude ash
Moisture> 14%
Moisture
Ash insoluble in HCl Max. 2%
2.2.2
Flaked barley.
Product obtained by steaming and rolling husked barley Hordeum vulgare L.
Crude fibre
Starch
Moisture> 14%
Moisture
Ash insoluble in HCl Max. 0.5%
2.2.3
Barley feed.
By-product of the processing of screened and husked barley into pearl barley or semolina or sifted barley meal.
Crude fibre
Starch
Crude ash
Moisture>14%
Moisture
Ash insoluble in HCl Max. 2%
2.2.4
Flaked Maize.
Product obtained by steaming and rolling maize.
Crude fibre
Starch
Moisture> 14%
Moisture
Ash insoluble in HCl Max. 0.5%
2.2.5
Pea middlings (pea forage meal).
By-product obtained during the manufacture of pea-meal Pisum sativum L. It consists principally of particles of endosperm, and to a lesser extent, of skins.
Crude fibre Crude oil and fat
Crude protein Moisture> 14%
Moisture
Ash insoluble in HCl Max. 2%
2.2.6
Flaked potatoes.
Product obtained by drying potatoes, Solanum tuberosum L., whether or not peeled, which have been steamed or boiled and crushed.
Crude fibre
Starch
Moisture> 14%
Moisture
Ash insoluble in HCl Max. 1.7%
2.3
By-products of maize milling.
2.3.1
Maize feed meal.
By-product of the manufacture of flour or semolina from maize.
Starch
Crude protein
Crude oil and fat Crude fibre
Moisture> 14%
Crude ash Moisture
Ash insoluble in HCl Max. 2%
2.3.2
Maize bran.
By-product of the manufacture of flour or semolina from maize. It consists principally of outer skins and maize germ, with some endosperm particles.
Crude fibre
Crude protein
Crude oil and fat
Moisture> 14%
Crude ash
Moisture
Ash insoluble in HCl Max. 2%
2.3.3
Maize germ and bran.
By-product of the manufacture of maize flour, maize semolina or of maize starch consisting of non-extracted germ, maize bran and some fragments of endosperm.
Crude oil and fat Crude protein
Crude fibre
Moisture> 13%
Crude ash
Starch
Moisture
Ash insoluble in HCl Max. 2%
2.4
Product and by-products of rice milling
2.4.1
Ground fodder rice.
Product obtained by grinding fodder rice consisting either of green, chalky or unripe grains, sifted out during the milling of husked rice, or of normal husked grains which are yellow or spotted.
Starch
Moisture> 14%
Crude fibre
Crude ash
Crude oil and fat
Crude protein Moisture
Ash insoluble in HCl Max. 1%
2.4.2
Broken rice.
By-product of the preparation of polished or glazed rice. It consists principally of under-sized or broken grains.
Starch
Moisture> 14%
Ash insoluble in HCl Max. 1%
2.4.3
Rice bran (brown)
By-product of the first polishing of husked rice. It consists of silvery skins, particles of the aleurone layer, endosperm and germ.
Crude protein
Crude fibre
Crde oil and fat
Crude ash
Moisture> 12%
Ash insoluble in HCl.
Moisture
Ash insoluble in HCl Max. 1.7%
2.4.4
Rice bran (white).
By-product of the second polishing of husked rice. It consists principally of particles of endosperm, of the aleurone layer and of germ.
Crude protein
Crude fibre
Crude oil and fat
Crude ash
Moisture> 12%
Ash insoluble in HCl.
Moisture
Ash insoluble in HCl Max. 0.6%
2.5
Products and by-products of the starch industry.
2.5.1
Maize starch
Virtually pure maize starch.
Starch
Moisture> 14%
Crude ash
Moisture
Ash insoluble in HCl Max. 2%
2.5.2
Puffed maize starch.
Virtually pure maize starch, greatly expanded by appropriate heat treatment.
Starch
Moisture> 10%
Crude ash Moisture
Ash insoluble in HCl Max. 0.5%
2.5.3
Pre-gelatinized partially hydrolyzed maize starch.
Virtually pure maize starch, largely pre-gelatinized and partially hydrolyzed.
Starch
Reducing sugars expressed as glucose Moisture> 10%
Crude ash Moisture
Ash insoluble in HCl Max. 0.5%
2.5.4
Maize gluten.
Dried by-product of the manufacture of maize starch. It consists principally of gluten obtained during the separation of the starch.
Crude protein
Crude oil and fat Moisture> 13%
Crude fibre
Crude ash Xanthophyll Moisture
Ash insoluble in HCl Max. 0.5%
2.5.5
Maize gluten feed
Dried by-products of the manufacture of maize starch. It is composed of bran and of a smaller quantity of gluten. Dried residues of the steeping liquors, and germ, from which the oil has been removed, have been added.
Crude protein
Crude fibre
Crude ash
Crude oil and fat Moisture> 13%
Moisture
Ash insoluble in HCl Max. 2%
2.5.6
Rice starch
Virtually pure rice starch
Starch
Moisture> 14%
Crude ash
Moisture
Ash insoluble in HCl Max. 0.5%
2.5.7
Puffed rice starch
Virtually pure rice starch, greatly expanded by appropriate heat treatment.
Starch
Moisture> 10%
Crude ash
Moisture
Ash insoluble in HCl Max. 0.5%
2.5.8
Rice gluten
Dried by-product of the manufacture of rice starch, consisting mainly of gluten.
Crude protein Moisture> 13%
Crude fibre
Crude ash
Crude oil & fat Moisture
Ash insoluble in HCl Max. 0.5%
2.5.9
Sorghum gluten feed
Dried by-product of the manufacture of sorghum starch Sorghum bicolor (L.) Moench s. 1. It consists of bran and a smaller quantity of gluten. Dried residues of the steeping liquors and the germ may be added.
Crude protein
Crude oil & fat
Crude fibre
Crude ash
Moisture> 13%
Moisture
Ash insoluble in HCl Max. 2%
2.5.10
Wheat starch
Virtually pure wheat starch
Starch
Moisture> 14%
Crude ash
Moisture
Ash insoluble in HCl Max. 0.5%
2.5.11
Puffed wheat starch
Virtually pure wheat starch greatly expanded by appropriate heat treatment.
Starch
Moisture> 10%
Crude ash Moisture
Ash insoluble in HCl Max. 0.5%
2.5.12
Pre-gelatinized, partially hydrolyzed wheat starch
Virtually pure wheat starch, largely pregelatinized and partially hydrolyzed.
Starch Reducing sugar expressed as glucose
Moisture> 10%
Crude ash Moisture
Ash insoluble in HCl Max. 0.5%
2.5.13
Wheat gluten
Dried by-product of the manufacture of wheat starch. It consists principally of gluten obtained during the separation of starch.
Crude protein Moisture> 12%
Crude ash Moisture
Ash insoluble in HCl Max. 0.5%
2.5.14
Manioc starch
Virtually pure starch obtained from manioc roofs Manihot esculenta Crantz.
Starch
Moisture> 15%
Crude ash Moisture
Ash insoluble in HCl Max. 0.5%
2.5.15
Puffed manioc starch
Starch obtained from manioc roots, greatly expanded by appropriate heat treatment.
Starch
Moisture> 10%
Crude ash Moisture
Ash insoluble in HCl Max. 0.5%
2.5.16
Potato starch
Virtually pure potato starch.
Starch
Moisture> 14%
Crude ash Moisture
Ash insoluble in HCl Max. 0.5%
2.5.17
Pre-gelatinized potato starch
Virtually pure potato starch, greatly expanded by appropriate heat treatment.
Starch
Moisture> 10%
Crude ash Moisture
Ash insoluble in HCl Max. 0.5%
2.5.18
Pre-gelatinized partially hydrolyzed potato starch
Virtually pure potato starch, greatly expanded and partially hydrolyzed.
Starch Reducing sugars expressed as glucose
Moisture> 10%
Crude ash Moisture
Ash insoluble in HCl Max. 0.5%
2.5.19
Potato protein
Dried by-product of starch manufacture composed mainly of protein substances obtained by the separation of starch.
Crude protein Moisture> 14%
Crude ash
Crude oil & fat Crude fibre Moisture
Ash insoluble in HCl Max. 0.5%
2.5.20
Dried potato pulp
Dried By-product of the manufacture of potato starch.
Starch
Moisture> 14%
Crude ash
Crude oil & fat Crude fibre Moisture
Ash insoluble in HCl Max. 2%
2.5.21
Dextrose (glucose).
Product of the saccharification of starch consisting of purified, crystallized glucose(with or without water of crystallization).
Glucose
Moisture> 10%
Moisture
Ash insoluble in HCl Max. 2%
2.5.22
Dextrose Molasses.
By-product obtained during the crystallization of dextrose.
Reducing sugars, expressed as glucose.
Crude ash
Moisture
Ash insoluble in HCl Max. 2%
2.6
Products and by-products of sugar manufacture
2.6.1
Sugar (sucrose).
Beet or cane sugar in solid form.
Sucrose
Crude ash
Ash insoluble in HCl Max. 2%
2.6.2
Dried sugar beet slices.
Product obtained by drying slices of washed sugar beet Beta vulgaris L., ssp. vulgaris var. altissima Doell.
Total sugar, expressed as sucrose Crude ash
Moisture> 13%
Moisture
Ash insoluble in HCl Max. 2%
2.6.3
Dried partially extracted sugar beet
Product obtained by drying washed sugar beet slices.
Total sugar, expressed as sucrose Crude ash
Moisture> 13%
Moisture
Ash insoluble in HCl Max. 2%
2.6.4
Dried plain sugar beet pulp
By-product of the manufacture of sugar, consisting of pulped and dried sugar beet slices.
Crude fibre
Ash insoluble in HCl Max. 3.5%
2.6.5
Sugar beet molasses.
By-product consisting of the syrupy residue collected during the manufacture or refining of beet sugar.
Total sugar, expressed as sucrose
Ash insoluble in HCl Max. 2%
2.6.6
Sugar cane molasses.
By-product consisting of the syrupy residue collected during the manufacture of refining of sugar from sugar cane Saccharum officinarum L.
Total sugar, expressed as sucrose
Ash insoluble in HCl Max. 2%
2.6.7
Wet sugar beet pulp.
By-product consisting of the undried residue resulting from the extraction of sugar from sugar beet.
Crude protein Crude oil
Crude fibre
Crude ash
Moisture
Ash insoluble in HCl Max. 2%
2.7
Products and by-products of malting, brewing distilling and fruit processing; dried feed yeasts
2.7.1
Barley malt culms.
By-product of malting, consisting of dried rootlets and shoots of germinated barley.
Crude protein
Crude fibre
Crude ash
Moisture> 12.5%
Moisture
Ash insoluble in HCl Max. 2%
2.7.2
Dried yeasts.
Yeasts, whether or not mixed, belonging to the families Saccharomycetaceae, Endo-mycetaceae and Cryptococcaceae, cultivated on the following substrates: beet or core juice or molasses, distillers' or yeast-makers' wash, whey, cereals and products derived from their processing, solutions from the hydrolysis of fibrous material, the cells of which have been killed by drying.
Crude protein
Crude ash
Moisture> 10%
Ash insoluble in HCl
Moisture
Ash insoluble in HCl Max. 1.1%
2.7.3
Dried brewers' grains.
By-product of brewing obtained by drying residues of malted and un-malted cereals and other starchy matter.
Crude protein
Crude fibre
Moisture> 12.5%
Moisture
Ash insoluble in HCl Max. 2%
2.7.4
Dried distillers' grains.
By-product of distilling obtained by drying residues of fermented cereals or other starchy matter.
Crude protein
Crude fibre
Moisture> 12.5%
Moisture
Ash insoluble in HCl Max. 2%
2.7.5
Dehydrated citrus pulp.
By-product obtained during the manufacture of citrus juice.
Crude fibre
Moisture> 13%
Moisture
Ash insoluble in HCl Max. 2%
2.7.6
Wet brewers grains.
By-product of brewing consisting of the undried residues of malted and unmalted cereals and other starchy matter.
Crude protein Crude oil
Crude fibre
Crude ash
Moisture
Ash insoluble in HCl Max. 2%.
2.7.7
Wet distillers grains.
By-product of distilling consisting of undried residues of fermented cereals or other starchy matter.
Crude protein Crude oil
Crude fibre
Crude ash Moisture
Ash insoluble in HCl Max. 2%
2.8
Artificially dried agricultural products
2.8.1
Grass meal.
Product obtained by artificially drying and possibly pre-drying young forage plants, the enzymes which activate oxidation being rendered virtually inactive by the drying.
Crude protein
Crude fibre
Crude ash
Moisture> 12%
Crude oil and fat Carotene Ash insoluble in HCl Moisture
Ash insoluble in HCl Max. 3.4%
2.8.2
Lucerne meal.
Product obtained by artificially drying and possibly pre-drying lucerne Medicago sativa L. and Medicago varia Martyn, the enzymes which active oxidation being rendered virtually inactive by the drying. This product may contain approximately 20% of grass or clover artificially dried and possibly pre-dried at the same time as the lucerne.
Crude protein
Crude ash
Crude fibre
Moisture> 12%
Crude oil and fat Carotene
Ash insoluble in HCl Moisture
Ash insoluble in HCl Max. 3.4%
2.8.3
Clover meal.
Product obtained by artificially drying and possible pre-drying young clover Trifolium spp. the enzymes which activate oxidation being rendered virtually inactive by the drying. This product may contain approximately 20% of grass or lucerne artifically dried and possibly pre-dried at the same time as the clover.
Crude protein
Crude ash
Crude fibre
Moisture> 12%
Crude oil and fat Carotene Ash insoluble in HCl Moisture
Ash insoluble in HCl Max. 3.4%
2.8.4
Dried tops and leaves of sugar beet.
Product obtained by artificially drying tops and leaves of sugar beet, washed, whether or not chopped.
Crude protein
Crude fibre
Total sugar, expressed as sucrose
Moisture> 12%
Ash insoluble in HCl Moisture
Ash insoluble in HCl Max. 4%
2.8.5
Jerusalem artichoke chips or Jerusalem artichoke meal.
Product obtained by crushing or grinding dried, cleaned tubers of Jerusalem artichokes Helianthus tuberosus L.
Inulin
Crude fibre
Moisture> 13%
Crude ash
Crude oil and fat Crude protein Moisture
Ash insoluble in HCl Max. 2%
2.8.6
Sweet potato chips or sweet potato meal.
Product obtained by crushing or grinding dried, cleaned tubers of sweet potato Ipomoea batatas (L.) Poir.
Starch
Moisture> 13%
Crude ash
Crude fibre
Crude oil and fat
Crude protein
Moisture
Ash insoluble in HCl Max. 2%
2.8.7
Manioc meal or manioc flakes or manioc roots.
Dried and, if necessary, washed and peeled manioc roots; also products obtained by crushing and grinding.
Starch
Crude ash
Moisture> 13%
Crude fibre
Crude oil and fat
Crude protein
Moisture
Ash insoluble in HCl Max. 3.3%
2.8.8
Manioc meal type 55 or manioc flakes type 55 or manioc roots type 55.
Unpeeled manioc roots, dried and, if necessary, washed; also products obtained by crushing and grinding.
Starch
Crude ash
Moisture> 13%
Crude fibre
Crude oil and fat
Crude protein
Moisture
Ash insoluble in HCl Max. 4%
2.8.9
Dried manioc pulp
Waste from the manufacture of manioc starch, which has been dried and ground.
Starch
Crude ash
Moisture> 13%
Crude fibre
Crude oil and fat
Crude protein
Moisture
Ash insoluble in HCl Max. 2.3%
2.9
Other products of vegetable origin
2.9.1
Crushed locust beans
Product obtained by crushing the dried, stoned fruit of the carob tree Ceratonia siliqua L.
Total sugar, expressed as sucrose
Moisture> 14%
Crude ash
Moisture
Ash insoluble in HCl Max. 2%
2.9.2
Vegetable fat or vegetable oil
Product composed of fat or oil of vegetable origin
Matter insoluble in light
petroleum> 1.5%
Acid lndex> 12 Moisture> 1%
Matter insoluble in light petroleum
Ash insoluble in HCl Max. 2%
2.9.3
Barley
Whole grain of cultivated varieties of barley
Hectolitre weight Moisture> 15%
Crude fibre
Crude protein Crude oil
Crude ash
Lysine
Methionine Cystine
Starch Moisture
Ash insoluble in HCl Max. 2%
2.9.4
Processed barley other than flaked barley(2.2.2) (indicate type of process in name)
Product obtained by the processing of whole barley
Crude fibre
Moisture> 14%
Crude protein Crude oil
Crude ash
Lysine
Methionine Cystine
Starch
Moisture
Ash insoluble in HCl Max. 2%
2.9.5
Oats
Whole grain of cultivated varieties of oats.
Hectolitre weight Moisture> 15%
Crude fibre
Crude protein Crude oil
Crude ash
Lysine
Methionine Cystine
Starch
Moisture
Ash insoluble in HCl Max. 2%
2.9.6
Processed oats (Indicate type of process in name)
Product obtained by the processing of whole oats
Crude fibre
Moisture> 14%
Crude protein Crude oil
Crude ash
Starch
Lysine
Methionine
Cystine Moisture
Ash insoluble in HCl Max. 2%
2.9.7
Wheat
Whole grain of cultivated varieties of wheat
Hectolitre weight Moisture>15%
Crude fibre
Crude protein Crude oil
Crude ash
Starch
Lysine
Methionine Cystine Moisture
Ash insoluble in HCl Max. 2%
2.9.8
Processed wheat(Indicate type of process in name)
Product obtained by the processing of whole wheat.
Crude fibre
Moisture> 14%
Crude protein Crude oil
Crude ash
Starch
Lysine
Methionine
Cystine Moisture
Ash insoluble in HCl Max. 2%
2.9.9
Maize
Whole grain of cultivated varieties of maize
Hectolitre weight Moisture> 14%
Crude fibre
Crude protein Crude oil
Crude ash
Starch
Lysine
Methionine
Cystine Moisture
Ash insoluble in HCl Max. 2%
2.9.10
Processed maize other than flaked maize (2.2.4) (Indicate type of process in name)
Product obtained by the processing of whole maize.
Moisture> 13%
Crude fibre
Crude protein Crude oil
Crude ash
Starch
Lysine
Methionine
Cystine Moisture
Ash insoluble in HCl Max. 2%
2.9.11
Milo
Whole grain of cultivated varieties of milo.
Hectolitre weight Moisture> 14%
Crude fibre
Crude protein Crude oil
Crude ash
Starch
Lysine
Methionine
Cystine Moisture
Ash insoluble in HCl Max. 2%
2.9.12
Processed milo (Indicate type of process in name)
Product obtained by the processing of whole milo.
Moisture> 13%
Crude fibre
Crude protein Crude oil
Crude ash
Starch
Lysine
Methionine
Cystine Moisture
Ash insoluble in HCl Max. 2%
2.9.13
Peas
Seeds of the cultivated varieties of peas
Crude protein Moisture> 14%
Crude oil
Crude fibre
Crude ash
Lysine
Methionine
Cystine Moisture
Ash insoluble in HCl Max. 2%
2.9.14
Processed peas (Indicate type of process in name)
Product obtained by the processing of whole peas
Crude protein Moisture> 14%
Crude oil
Crude fibre
Crude ash
Lysine
Methionine
Cystine Moisture
Ash insoluble in HCl Max. 2%
2.9.15
Beans
Seeds of the cultivated varieties of beans
Crude protein Moisture> 14%
Crude oil
Crude fibre
Crude ash
Lysine
Methionine
Cystine Moisture
Ash insoluble in HCl Max. 2%
2.9.16
Processed beans (Indicate type of process in name)
Product obtained by the processing of whole beans.
Crude protein Moisture> 14%
Crude oil
Crude fibre
Crude ash
Lysine
Methionine
Cystine
Moisture
Ash insoluble in HCl Max. 2%
2.9.17
Seaweed meal
Product obtained by the processing of seaweed.
Crude ash
Moisture> 14%
Crude protein Crude oil
Crude fibre Calcium Phosphorus Moisture
Ash insoluble in HCl Max. 2%
2.9.18
Rape seed
Seeds of colza Brassica napus L. ssp. oleifera (Metzg.) Sinsk., of Indian sarson Brassica napus L. var. glauca (Roxb.) O.E Schulz and of rape Brassica campestvis L. ssp. oleifera (Metzg.) Sinsk
Crude protein
Crude oil and fat Crude fibre
Crude ash
Moisture> 9%
Moisture
Ash insoluble in HCl Max. 2%
2.9.19
Heat processed soya beans
Product obtained from heating whole soya beans without removing any of the component parts.
Crude Protein
Crude oil and fat Crude fibre
Crude ash
Moisture> 12%
Moisture
Ash insoluble in HCl Max. 2%
3.
Products of Animal Origin
3.1
Milk products
3.1.1
'Spray' skimmed-milk powder, 'hatmaker' or 'roller' skimmed-milk powder
Product obtained by drying skimmed milk either by vaporization in a current of hot air ('spray' skimmed milk powder) or by drying over cylinders ('hatmaker' or 'roller' skimmed milk).
Crude protein
Lactose
Crude ash
Moisture> 5%
Crude oil & fat
Moisture
Ash insoluble in HCl Max. 0.5%
3.1.2
Powdered buttermilk
Product obtained by drying buttermilk either by vaporization in a current of hot air ('spray' powdered buttermilk) or by drying over cylinders ('hatmaker' or 'roller' powdered buttermilk).
Crude protein
Crude oil & fat Lactose
Crude ash
Moisture> 6%
Moisture
Ash insoluble in HCl Max. 0.5%
3.1.3
Powdered whey or whey crumbs
Products obtained by drying whey.
Crude protein
Lactose
Crude ash
Sodium
Moisture> 8%
Crude oil and fat Chlorides, expressed as NaCl
Moisture
Ash insoluble in HCl Max. 0.5%
3.1.4
Low-sugar powdered whey
Product obtained by drying whey from which the lactose has been partly extracted
Lactose
Crude protein
Crude ash
Sodium
Moisture> 8%
Crude oil and fat
Chlorides, expressed as NaCl
Moisture
Ash insoluble in HCl Max. 0.5%
3.1.5
Powdered whey protein
Powdered milk albumin
Products obtained by drying the protein compounds extracted from whey or milk by chemical or physical treatment.
Crude protein
Crude ash
Moisture> 8%
Crude oil and fat
Moisture
Ash insoluble in HCl Max. 0.5%
3.1.6
Skim milk
Residue obtained following the removal of fat from whole milk.
Total solids
Crude protein Lactose
Ash
Ash insoluble in HCl Max. 2%
3.1.7
Whey
By-product obtained during the manufacture of cheese.
Total solids
Crude protein Lactose
Ash
Sodium
Ash insoluble in HCl Max. 2%
3.1.8
Concentrated whey
Product obtained by removing part of the moisture from whey.
Total solids
Crude Protein
Lactose
Ash
Sodium
Ash insoluble in HCl Max. 2%
3.2
Products processed from land animals
3.2.1
Blood Meal
Product obtained by drying the blood of slaughtered animals and poultry. This product should be substantially free of foreign matter.
Crude Protein
Moisture> 10%
Crude ash Moisture>10%
Ash insoluble in HCl Max. 2%
3.2.2
Meat and bone meal
Product obtained by drying and grinding meat pieces containing a high proportion of bone from warm-blooded land animals. The product should be substantially free of hair, bristle, feathers, horn, hoof, skin and blood, and of the contents of the stomach and viscera. It must be technically free from organic solvents.
Crude protein
Crude oil & fat
Total phosphorus Crude ash
Moisture> 10%
Chlorides, expressed as NaCl Methionine
Lysine
Volatile nitrogenous bases
Moisture
Ash insoluble in HCl Max. 2%
3.2.3
Bone Meal
Product obtained by drying and grinding bone, with the fat largely removed, from warm-blooded land animals. The product should be substantially free of hair, bristle, feathers, horn, hoof, skin and blood, and of the contents of the stomach and viscera. It should also be free of splinters and may not contain bone fragments with rough surfaces or jagged edges. It must be technically free from organic solvents.
Crude protein
Total phosphorus
Crude ash
Crude oil & fat Moisture> 10%
Moisture
Ash insoluble in HCl Max. 2%
3.2.4
Meat meal. Products with a fat content of more than 11% should be described as 'rich in fat'
Product obtained by drying and grinding carcases and parts of carcases of warm-blooded land animals, if need be, with the fat removed by an appropriate process. It should be virtually free of hair, bristle, feathers, horn, hoof and skin, and the contents of the stomach and viscera. It shall be technically free of organic solvents.
Crude protein
Crude oil & fat
Total phosphorus Moisture> 10%
Chlorides, expressed as NaCl Ash insoluble in HCl
Methionine
Lysine
Volatile nitrogenous bases Moisture
Ash insoluble in HCl Max. 2.2%
3.2.5
Greaves
Product derived from residues of the manufacture of tallow and other fats of animal origin. It shall be technically free of organic solvents.
Crude protein
Crude ash
Moisture> 10%
Chlorides, expressed as NaCl Crude oil & fat Moisture
Ash insoluble in HCl Max. 0.5%
3.2.6
Dried waste from poultry slaughter. Products with a fat content of more than 12% should be described as 'rich in fat'.
Product obtained by drying and grinding waste from slaughtered poultry; it should be substantially free of feathers.
Crude protein
Crude oil & fat
Crude ash
Moisture> 10%
Chlorides, expressed as NaCl
Moisture
Ash insoluble in HCl Max. 3.3%
3.2.7
Hydrolyzed feather meal.
Product obtained by hydrolyzing, drying and grinding poultry feathers.
Crude protein
Moisture> 11%
Ash insoluble in HCl
Moisture
Ash insoluble in HCl Max 3.4%
3.2.8
Animal fat
Product composed of fat processed from warm-blooded land animals or from parts thereof. It shall be technically free of organic solvents.
Acid Index> 30 Matter insoluble in light
petroleum> 1.5% Moisture> 1%
Acid index
Ash insoluble in HCl Max. 2%
3.3
Products derived from fish or other marine animals.
3.3.1
Fish meal (products whose chloride content expressed as NaCl is less than 2% may be referred to as 'low in salt')
Product obtained by drying and grinding whole fish, or parts thereof, of various species. Concentrated press liquid may be added.
Crude protein
Crude oil & fat Chlorides, expressed as NaCl
Total phosphorus Calcium carbonate Moisture> 10%
Moisture
Ash insoluble in HCl Max. 2.2%
3.3.2
Cod liver oil
Oil obtained from fresh livers of fish of the cod family(Gadidae)
Vitamin A
Acid index> 6 Matter insoluble in light
petroleum> 0.05%
Moisture> 0.15%
Acid index0.15%
Ash insoluble in HClMax. 2%
4.
MINERAL SUBSTANCES
4.1
Calcium carbonate (the nature of the product(column 3) should be indicated in the name)
Precipitated calcium carbonate, ground limestone, prepared chalk, granulated chalk, ground oyster or mussel shells.
Calcium
Ash insoluble in HCl
Ash insoluble in HCl Max. 5%
4.2
Calcium and magnesium carbonate
Natural mixture of calcium carbonate and magnesium carbonate.
Calcium
Magnesium
Ash insoluble in HCl Max. 2%
4.3
Calcareous marine algae (Maerl)
Product of natural origin obtained from calcareous algae, ground or granulated.
Calcium
Ash insoluble in HCl
Ash insoluble in HCl Max. 5%
4.4
Magnesium oxide
Technically pure magnesium oxide MgO
Magnesium
Ash insoluble in HCl Max. 2%
4.5
Kieserite
Natural magnesium sulphateMgSO
4
H
2
O
Magnesium
Ash insoluble in HCl Max. 2%
4.6
Calcium monohydrogen phosphate (dicalcium phosphate) (the manufacturing process may be indicated in the name)
Technically pure calcium monohydrogen phosphate (dicalcium phosphate).
Total phosphorus Chlorides, expressed as NaCl
Calcium
Ash insoluble in HCl Max. 2%
4.7
Defluorinated natural phosphate
Product obtained by grinding natural phosphates, purified and defluorinated to a greater or lesser degree.
Total phosphorus Calcium
Ash insoluble in HCl Max. 2%
4.8
De-gelatinized bone meal
De-gelatinized, sterilized, ground bones from which the fat has been removed.
Total phosphorus
Calcium
Moisture> 10%
Moisture
Ash insoluble in HCl Max. 2%
4.9
Calcium bis- (dihydrogen phosphate) (monocalcium phosphate)
Product consisting principally of technically pure calcium bis- (dihydrogen phosphate) (monocalcium phosphate)
Total phosphorus
Calcium
Ash insoluble in HCl Max. 2%
4.10
Ammonium dihydrogen phosphate (monoammonium phosphate)
Product consisting mainly of technically pure ammonium dihydrogen phosphate.
Total phosphorus Nitrogen
Ash insoluble in HCl Max. 2%
4.11
Salt
Technically pure sodium chloride (NaCl)
Chlorides, expressed as NaCl
Ash insoluble in HCl Max. 2%
4.12
Other mineral substances except trace elements Common name to be used.
Calcium> 5% Phosphorus> 5% Magnesium> 5% Sodium> 5%
Calcium
Ash insoluble in HCl Max. 2%
5
OTHER STRAIGHT FEEDINGSTUFFS
5.1
Common name to be used.
All feed ingredients not listed in Part I of this Schedule except those listed in Part II.
Crude protein>5% Crude oil &
fat> 2.5%
Crude fibre> 5% Crude ash> 4% Starch> 15% Moisture> 14%
Crude protein
Ash insoluble in HCl Max. 2%
PART II
PROHIBITED STRAIGHT FEEDINGSTUFFS
Bagasse including molassed bagasse
Cocoa shells
Corozo nut meal
Dried poultry manure
Grape by-products from wine manufacture (grape pulp, grape follicle meal etc.)
Grass seed cleanings
Hoof or horn meal
Rice hulls
Shea nut meal or karite nut meal
Sal seed meal.
PART III
COMPOUND FEEDINGSTUFFS
(Contents of analytical constituents to be declared as a percentage of the weight of the compound feedingstuff as such)
(1)
(2)
(3)
Type of compound feedingstuffs
Compulsory declaration (Regulation 8(1) (b) (vi))
Optional declarations
(Regulation 9(2) (d))
Compound
—crude protein
—soluble protein
feedingstuffs with
—crude oils and fats
—starch
the exception of
—crude fibre
—total sugar, expressed as
wholegrain mixes,
—moisture where it
sucrose
mineral and
equals or
—total sugar plus starch
molassed
exceeds 14%
—moisture
where the levelsare less than those specified in column 2
feedingstuffs and
—crude ash
—Calcium
compound
—calcium, where it
—magnesium
feedingstuffs for pet
equals or
—sodium
animals
exceeds 5%
—phosphorus-
—magnesium,
where it equals or
exceeds 0.4%
—sodium, where it
—lysine
equals or exceeds
—methionine
0.5%
—cystine
—phosphorus, where it equals or
exceeds 1%
—lysine, in compound
feedingstuffs for swine
—methionine in compound
feedingstuffs for poultry
—cystine in compound feedingstuffs for poultry
Mineral feedingstuffs
—calcium
—crude ash
—phosphorus
—crude protein
—sodium
—crude oils and fats
—magnesium
—crude fibre
—moisture content if it equals or exceeds 5% in mineral feedingstuffs containing no organic substances or 10% in mineral feedingstuffs containing organic substances
—moisture where the levels are less than those specified in column 2 opposite
—lysine, in mineral feedingstuffs for pigs
—methionine in mineral feedingstuffs for poultry
—cystine in mineral feedingstuffs for poultry
Molassed feedingstuffs
—crude protein
—soluble protein
—crude oils and fats
—starch
—crude fibre
—Total sugar plus starch
—moisture, where it equals or exceeds 14%
—moisture, where it is less than 14%
—crude ash
—calcium
—total sugar expressed as sucrose
—magnesium
—sodium
—phosphorus
—lysine
—methionine
—cystine
in feeds for pigs, poultry or preruminating ruminants
Compound feedingstuffs for cats and dogs
—Crude protein
—crude oils and fats
—calcium
—sodium
—crude fibre
—phosphorus
—crude ash
—moisture
Compound feedingstuffs for pet animals other than cats and dogs
—moisture
—crude protein
—crude oils and fats
—crude fibre
—crude ash
—calcium
—sodium
—phosphorus
SECOND SCHEDULE
Fees Referred to in Regulation 11 (2) (b).
IR£
Moisture
4
Crude protein
6
Soluble Protein
15
Volatile nitrogenous bases
15
Amino acids
25 for one
amino
acid plus
2 for each
addition
al amino
acid
Crude oils and fats
6
Acid index
10
Matter insoluble in light petroleum
10
Crude fibre
6
Starch
10
Total sugars
10
Reducing sugars
10
Lactose
10
Glucose
10
Total solids
5
Skim milk powder
15
Crude ash
3
Ash soluble in HCl
5
Calcium carbonate
10
Chlorides expressed as NaCl
5
Calcium
10
Phosphorus
10
Sodium
10
Potassium
10
Magnesium
10
Bushel Weight
4
Vitamin A
15
THIRD SCHEDULE
PART I
I, AUSTIN DEASY, Minister for Agriculture, in exercise of the powers conferred on me by
section 3
of the
European Communities Act, 1972
(No. 27 of 1972), and for the purpose of giving effect to Council Directive No. 77/101/EEC of 23 November, 1976
1
as amended in the manner specified in relation thereto in Regulation 2 of the following regulations, and Council Directive No. 79/373/EEC of 2 April, 1979
2
as amended in the manner specified in relation thereto in the said Regulation 2, hereby make the following regulations
1
O.J. No. L32/1, 3 February 1977.
2
O.J. No. L86/30, 6 April 1979.