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GB 5413.3-2010: National food safety standard -- Determination of fat in foods for infants and young children, milk and milk products
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GB 5413.3-2010
GB
NATIONAL STANDARD
OF THE PEOPLE’S REPUBLIC OF CHINA
National food safety standard
Determination of fat in foods for infant and young
children, milk and milk products
ISSUED ON. MARCH 26, 2010
IMPLEMENTED ON. JUNE 1, 2010
Issued by. Ministry of Health of the People's Republic of China
Table of Contents
Preface ... 3 
1 Scope ... 4 
2 Normative documents ... 4 
Method 1 ... 4 
3 Principle ... 4 
4 Reagents and materials ... 4 
5 Instruments and apparatuses ... 5 
6 Test procedure ... 5 
7 Results calculation ... 9 
8 Precision ... 9 
9 Others ... 9 
Method 2 ... 10 
10 Principle ... 10 
11 Reagent and materials... 10 
12 Instruments and apparatuses ... 11 
13 Test procedure ... 11 
14 Precision ... 11 
Appendix A ... 12 
Preface
This standard replaces the determination of fat in GB/T 5009.46-2003 “Method of
analysis of hygienic standard of milk and milk products”; the determination of fat in
GB/T 5409-85 “Analytical method for milk”; the determination of fat in GB/T 5416-85
“Analytical method for butter”; and GB/T 5413.3-1997 “Milk powder and formula foods
for infant and young children - Determination of fat”.
Appendix A is informative.
The previous editions to be replaced by this standard are.
- GB/T 5409-85;
- GB/T 5413.3-1997;
- GB/T 5416-85;
- GB/T 5009.46-1985, GB/T 5009.46-1996, and GB/T 5009.46-2003.
National food safety standard
Determination of fat in foods for infant and young
children, milk and milk products
1 Scope
This standard specifies the determination of fat in pasteurized milk, sterilized milk, raw
milk, fermented milk, modulation milk, milk powder, condensed milk, butter, diluted
butter, cheese, and infant formula foods.
Method 1 in this standard is applicable to determination of fat in pasteurized milk,
sterilized milk, raw milk, fermented milk, modulation milk, milk powder, condensed milk,
butter, diluted butter, cheese, and infant formula foods. Method 2 in this standard is
applicable to determination of fat in pasteurized milk, sterilized milk, and raw milk.
2 Normative documents
The normative documents are indispensable to this Standard. For dated references,
only the dated references apply to this Standard. For undated references, the latest
editions of the normative documents (including all corrigendum) to apply to this Standard.
Method 1
3 Principle
Use ethyl ether and petroleum ether to extract the alkaline hydrolyzate of the sample.
Distil or evaporate to remove the solvent. Determine the extract’s mass that is dissolved
in the solvent.
4 Reagents and materials
Unless otherwise specified, all reagents are analytically pure. Water is the Grade 3
specified in GB/T 6682.
4.1 Amylase. Enzyme activity ≥ 1.5 U/mg.
4.2 Ammonia (NH4OH). Mass fraction is about 25%.
Note. Higher concentration of ammonia may be used.
4.3 Ethanol (C2H5OH). Volume fraction ≥ 95%.
4.4 Ethyl ether (C4H10O). Does not contain peroxide or antioxidant; and meet the test
requirements.
4.5 Petroleum ether(CnH2n+2). Boiling range 30 - 60 °C.
4.6 Mixed-solvent. Mix ethyl ether (4.4) with same-volume’s petroleum ether (4.5).
Prepare before using.
4.7 Iodine solution (I2). About 0.1 mol/L.
4.8 Congo-red solution (C32H22N6Na2O6S2). Dissolve 1g of Congo-red into water; dilute
to 100ml.
Note. It may be selectively used. Congo-red can make the interface of solvent and water-phase to
be clear. Other solutions can also be used if it can colorize the water-phase and do not affect the
test result.
4.9 Hydrochloric acid (6 mol/L). Pipette 50 mL of hydrochloric acid to slowly pour into 40
mL of water. Fix the volume to 100 mL. Mix well.
5 Instruments and apparatuses
5.1 Analysis balance. Sensitivity is 0.1mg.
5.2 Centrifuge. It can be used for placement of liposuction bottle or tube. Rotating
speed is 500-600 r/min. It can generate 80-90g gravity field at the outer end of
liposuction bottle.
5.3 Drying Oven.
5.4 Water bath.
5.5 Liposuction bottle. With high quality cork stopper or other stoppers (e.g. silica gel
or polytetrafluoroethylene) which do not affect the solvent use. Firstly immerse the cork
stopper in ethyl ether; then put into the water with temperature of 60°C or above for
minimum 15min. Cool down before using. Immerse the cork stopper in water when it is
not in use. Change the immersing water daily.
Note. It may also liposuction tube (or bottle) with siphon. However, the operational steps are
different, see Appendix A. The lower-end of the internal long branch-pipe of the joint can be a
spoon-shaped.
6 Test procedure
6.1 Preparation of the fat-collection container (fat-collection bottle)
Put a few grains of zeolites in a dried fat-collection bottle. Place it a drying oven to dry
for 1 hour. Cool the fat-collection bottle to room temperature. Weigh to precision of 0.1
mg.
Note. The fat-collection bottle may be selected according to actual needs.
6.2 Blank test
Avoid splashing the solvent outside of the bottle. Place the fat-collection bottle into
drying oven at 102°C±2°C. Heat for 1 hour. Operate according to 6.3.1.12 and
6.3.1.13.
6.3.1.16 The difference between the mass from 6.3.1.13 AND mass from 6.3.1.15 is
the content of fat.
Note. Follow the description in Appendix A when selecting liposuction tube with a siphon or wash-
bottle accessory.
6.3.2 Milk powder and milk-based infant food
Weigh the well-mixed sample. High-fat milk powder, full cream milk powder, full cream
milk powder with sugar, and milk-based infant food. ~ 1 g (accurate to 0.0001 g);
skimmed milk powder, whey powder, and buttermilk powder. ~ 1.5 g (accurate to
0.0001 g).
6.3.2.1 Sample without starch
Add 10mL of water at temperature 65 ±5°C. Wash the sample into the small ball in
the liposuction bottle. Mix thoroughly till the sample is fully dispersed. Place it in
running water to cool down.
6.3.2.2 Sample with starch
Put the sample into liposuction bottle. Add 0.1g of amylase (4.1) and a magnetic
stirring rod . After mixing well, add 8 mL - 10mL distilled water at 45 °C. The solution
surface should not be too high. Stuffed with cork stopper. Under stirring state, place it
in the 65 °C water bath for 2 hours. Shake every 10 min. In order to verify if the starch
is fully hydrolyzed, it may add two drops of 0.1ml/L iodine solution (4.7). It indicates
fully hydrolyzation if there is no blue color occurred. Otherwise put the bottle back into
water bath, till there is no blue color observed. Cool down the liposuction bottle.
Follow the operations of 6.3.1.1 - 6.3.1.16.
6.3.3 Condensed milk
For skimmed condensed milk, full cream condensed milk and partially skimmed
condensed milk, weigh about 3 g – 5 g; for high fat condensed milk, weigh about 1.5 g
(accurate to 0.0001g). Use 10mL of distilled water to wash into the liposuction bottle’s
small-ball, in several times. Mix well.
Follow the operations of 6.3.1.1 - 6.3.1.16.
6.3.4 Butter and diluted butter
Dissolve the butter sample in warm water bath and mix thoroughly. Weigh about 0.5g
(accurate to 0.0001g) of sample. For diluted butter, weigh 1g into liposuction bottle.
Add 8mL - 10mL of distilled water at 45°C . Add 2 mL of ammonia to mix well.
Follow the operations of 6.3.1.1 - 6.3.1.16.
6.3.5 Cheese
To conduct the blank test, put 1g of anhydrous cream into the fat-collection bottle. If
necessary, add 1g of anhydrous cream in each 100ml of solvent; then re-distillation.
Use as quickly as possible after the re-distillation.
9.2 Blank test shall be in parallel with sample test, at the same time
For reagents containing non-volatile substances, it may be calibrated by the blank test
that is performed at the same time with the sample determination. The temperature
difference between the balance chamber and fat-collection bottle can affect the actual
mass of the extracts. Under ideal conditions (low blank value of reagent; temperatu...
GB 5413.3-2010
GB
NATIONAL STANDARD
OF THE PEOPLE’S REPUBLIC OF CHINA
National food safety standard
Determination of fat in foods for infant and young
children, milk and milk products
ISSUED ON. MARCH 26, 2010
IMPLEMENTED ON. JUNE 1, 2010
Issued by. Ministry of Health of the People's Republic of China
Table of Contents
Preface ... 3 
1 Scope ... 4 
2 Normative documents ... 4 
Method 1 ... 4 
3 Principle ... 4 
4 Reagents and materials ... 4 
5 Instruments and apparatuses ... 5 
6 Test procedure ... 5 
7 Results calculation ... 9 
8 Precision ... 9 
9 Others ... 9 
Method 2 ... 10 
10 Principle ... 10 
11 Reagent and materials... 10 
12 Instruments and apparatuses ... 11 
13 Test procedure ... 11 
14 Precision ... 11 
Appendix A ... 12 
Preface
This standard replaces the determination of fat in GB/T 5009.46-2003 “Method of
analysis of hygienic standard of milk and milk products”; the determination of fat in
GB/T 5409-85 “Analytical method for milk”; the determination of fat in GB/T 5416-85
“Analytical method for butter”; and GB/T 5413.3-1997 “Milk powder and formula foods
for infant and young children - Determination of fat”.
Appendix A is informative.
The previous editions to be replaced by this standard are.
- GB/T 5409-85;
- GB/T 5413.3-1997;
- GB/T 5416-85;
- GB/T 5009.46-1985, GB/T 5009.46-1996, and GB/T 5009.46-2003.
National food safety standard
Determination of fat in foods for infant and young
children, milk and milk products
1 Scope
This standard specifies the determination of fat in pasteurized milk, sterilized milk, raw
milk, fermented milk, modulation milk, milk powder, condensed milk, butter, diluted
butter, cheese, and infant formula foods.
Method 1 in this standard is applicable to determination of fat in pasteurized milk,
sterilized milk, raw milk, fermented milk, modulation milk, milk powder, condensed milk,
butter, diluted butter, cheese, and infant formula foods. Method 2 in this standard is
applicable to determination of fat in pasteurized milk, sterilized milk, and raw milk.
2 Normative documents
The normative documents are indispensable to this Standard. For dated references,
only the dated references apply to this Standard. For undated references, the latest
editions of the normative documents (including all corrigendum) to apply to this Standard.
Method 1
3 Principle
Use ethyl ether and petroleum ether to extract the alkaline hydrolyzate of the sample.
Distil or evaporate to remove the solvent. Determine the extract’s mass that is dissolved
in the solvent.
4 Reagents and materials
Unless otherwise specified, all reagents are analytically pure. Water is the Grade 3
specified in GB/T 6682.
4.1 Amylase. Enzyme activity ≥ 1.5 U/mg.
4.2 Ammonia (NH4OH). Mass fraction is about 25%.
Note. Higher concentration of ammonia may be used.
4.3 Ethanol (C2H5OH). Volume fraction ≥ 95%.
4.4 Ethyl ether (C4H10O). Does not contain peroxide or antioxidant; and meet the test
requirements.
4.5 Petroleum ether(CnH2n+2). Boiling range 30 - 60 °C.
4.6 Mixed-solvent. Mix ethyl ether (4.4) with same-volume’s petroleum ether (4.5).
Prepare before using.
4.7 Iodine solution (I2). About 0.1 mol/L.
4.8 Congo-red solution (C32H22N6Na2O6S2). Dissolve 1g of Congo-red into water; dilute
to 100ml.
Note. It may be selectively used. Congo-red can make the interface of solvent and water-phase to
be clear. Other solutions can also be used if it can colorize the water-phase and do not affect the
test result.
4.9 Hydrochloric acid (6 mol/L). Pipette 50 mL of hydrochloric acid to slowly pour into 40
mL of water. Fix the volume to 100 mL. Mix well.
5 Instruments and apparatuses
5.1 Analysis balance. Sensitivity is 0.1mg.
5.2 Centrifuge. It can be used for placement of liposuction bottle or tube. Rotating
speed is 500-600 r/min. It can generate 80-90g gravity field at the outer end of
liposuction bottle.
5.3 Drying Oven.
5.4 Water bath.
5.5 Liposuction bottle. With high quality cork stopper or other stoppers (e.g. silica gel
or polytetrafluoroethylene) which do not affect the solvent use. Firstly immerse the cork
stopper in ethyl ether; then put into the water with temperature of 60°C or above for
minimum 15min. Cool down before using. Immerse the cork stopper in water when it is
not in use. Change the immersing water daily.
Note. It may also liposuction tube (or bottle) with siphon. However, the operational steps are
different, see Appendix A. The lower-end of the internal long branch-pipe of the joint can be a
spoon-shaped.
6 Test procedure
6.1 Preparation of the fat-collection container (fat-collection bottle)
Put a few grains of zeolites in a dried fat-collection bottle. Place it a drying oven to dry
for 1 hour. Cool the fat-collection bottle to room temperature. Weigh to precision of 0.1
mg.
Note. The fat-collection bottle may be selected according to actual needs.
6.2 Blank test
Avoid splashing the solvent outside of the bottle. Place the fat-collection bottle into
drying oven at 102°C±2°C. Heat for 1 hour. Operate according to 6.3.1.12 and
6.3.1.13.
6.3.1.16 The difference between the mass from 6.3.1.13 AND mass from 6.3.1.15 is
the content of fat.
Note. Follow the description in Appendix A when selecting liposuction tube with a siphon or wash-
bottle accessory.
6.3.2 Milk powder and milk-based infant food
Weigh the well-mixed sample. High-fat milk powder, full cream milk powder, full cream
milk powder with sugar, and milk-based infant food. ~ 1 g (accurate to 0.0001 g);
skimmed milk powder, whey powder, and buttermilk powder. ~ 1.5 g (accurate to
0.0001 g).
6.3.2.1 Sample without starch
Add 10mL of water at temperature 65 ±5°C. Wash the sample into the small ball in
the liposuction bottle. Mix thoroughly till the sample is fully dispersed. Place it in
running water to cool down.
6.3.2.2 Sample with starch
Put the sample into liposuction bottle. Add 0.1g of amylase...
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