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GB 5009.128-2016 English PDF (GB5009.128-2016)

GB 5009.128-2016 English PDF (GB5009.128-2016)

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GB 5009.128-2016: National food safety standard -- Determination of cholesterin in foods

This Standard specifies the determination methods of cholesterol in foods. This Standard applies to the determination of cholesterol in foods. Method One Gas chromatography is applicable to the determination of cholesterol in meat and meat products, eggs and egg products, milk and dairy products and other animal foods and vegetable oils and fats. Method Two High performance liquid chromatography is applicable to the determination of cholesterol in meat and meat products, eggs and egg products, milk and dairy products and other animal foods. Method Three Colorimetric method is applicable to the determination of cholesterol in meat and meat products, eggs and egg products and other animal foods.
GB 5009.128-2016
GB
NATIONAL STANDARD OF
THE PEOPLE REPUBLIC OF CHINA
National food safety standard -
Determination of cholesterol in foods
ISSUED ON. DECEMBER 23, 2016
IMPLEMENTED ON. JUNE 23, 2017
Issued by. National Health and Family Planning Commission of the
People's Republic of China;
China Food and Drug Administration.
Table of Contents
Foreword ... 3
1 Scope ... 4
2 Principle... 4
3 Reagents and materials ... 4
4 Instruments and equipment ... 5
5 Analysis steps ... 5
6 Expression of analysis result ... 7
7 Precision... 8
8 Other ... 8
9 Principle... 8
10 Reagents and materials... 8
11 Instruments and equipment ... 9
12 Analysis steps ... 9
13 Expression of analysis result ... 11
14 Precision ... 12
15 Other ... 12
16 Principle... 12
17 Reagents and materials... 12
18 Instruments and equipment ... 14
19 Analysis steps ... 14
20 Expression of analysis result ... 15
21 Precision ... 15
22 Other ... 15
Annex A Cholesterol standard chromatogram ... 16
Foreword
This Standard replaces GB/T 5009.128-2003 Determination of cholesterol in foods, GB/T 22220-2008 Determination of cholesterol in foods - High-
performance liquid chromatography and GB/T 9695.24-2008 Meat and meat
products - Determination of cholesterol.
Compared with GB/T 5009.128-2003, the main changes in this Standard are as follows.
- modified the standard?€?s name to ?€?National food safety standard -
Determination of cholesterol in foods?€?;
- added gas chromatography as method one and high-performance liquid
chromatography as method two; modified colorimetric method as method
three;
- modified extraction solvent, the amount of ethanol and volume constant volume in pre-treatment method of gas chromatography in GB/T 9695.24-
2008.
National food safety standard -
Determination of cholesterol in foods
1 Scope
This Standard specifies the determination methods of cholesterol in foods. This Standard applies to the determination of cholesterol in foods. Method One Gas chromatography is applicable to the determination of cholesterol in meat and meat products, eggs and egg products, milk and dairy products and other animal foods and vegetable oils and fats. Method Two High performance liquid chromatography is applicable to the determination of cholesterol in meat and meat products, eggs and egg products, milk and dairy products and other animal foods. Method Three Colorimetric method is applicable to the
determination of cholesterol in meat and meat products, eggs and egg products and other animal foods.
Method One -- Gas chromatography
2 Principle
Saponify the sample by absolute ethanol - potassium hydroxide solution. Extract by petroleum ether and anhydrous ether mixture. Concentrate the extract to dry. After dissolving and setting volume of absolute ethanol, use gas chromatography to detect, external standard method to quantify.
3 Reagents and materials
Unless otherwise indicated, the reagents used in this method are of analytical grade, water is grade one water specified in GB/T 6682.
3.1 Reagents
3.1.1 methanol (CH3OH). chromatographic pure
3.1.2 anhydrous ethanol (C2H5OH)
3.1.3 petroleum ether. boiling range 30??C ~ 60??C
3.1.4 anhydrous ether (C4H10O).
Take 200 g of edible part of sample to homogenize. Place the sample in a sealed container to prevent deterioration and composition changes. The
sample shall be analyzed as soon as possible within 24 hours of
homogenization.
5.1.2 Vegetable oil, dairy products and other liquid samples
Take the uniform liquid sample after mixing into a sealed container to be tested. 5.2 Sample processing
5.2.1 Saponification
Weigh 0.25g ~ 10g of prepared sample (nearest to 0.001 g, cholesterol content of about 0.5mg ~ 5mg) in a 250-mL round-bottomed flask. Add 30 mL of
absolute ethanol, 10 mL of 60% potassium hydroxide solution. Well mix. Heat the sample at 100??C magnetic stirring heater for saponification reflux 1h. Shake from time to time so as to prevent sample sticking to the bottle wall. After the saponification ends, use 5 mL of absolute ethanol to rinse its interior from the condenser top. Remove the round-bottomed flask. Cool to room temperature with running water.
5.2.2 Extraction
Quantitatively transfer of all saponified liquid in a 250-mL separatory funnel. Use 30 mL of water to rinse the round-bottomed flask in 2~3 times. Merge the washing liquid into the separatory funnel. Then use 40 mL of petroleum ether - anhydrous ether mixture (1+1, volume ratio) to rinse the round-bottomed flask in 2~3 times. Then merge into the separatory funnel. Shake 2 min, place still and stratify. Transfer the water phase. Combine the three organic phases. Wash the extract to neutral with 100 mL of water each time. Swirl gently for the first time to prevent emulsification. The extract shall be dehydrated with about 10 g of anhydrous sodium sulfate to a 150-mL flask.
5.2.3 Concentration
Evaporate the extract in the flat-bottomed flask to near dryness under vacuum condition. Use absolute ethanol to dissolve and set volume to 5 mL. Determine with gas chromatograph.
The pretreatment of different sample requires blank test at the same time. 5.3 Determination
5.3.1 Instrument reference conditions
a) Chromatographic column. DB-5 flexible quartz capillary column, column length of 30 m, inner diameter of 0.32 mm, particle size of 0.25 ??m, or Take 200 g of edible part of sample. Use a meat grinder or homogenizer to homogenize the sample. Place the sample in a sealed container to prevent deterioration and composition changes. The sample shall be analyzed as soon as possible within 24 hours of homogenization
12.1.2 Dairy and other liquid samples
Take well mixed uniform liquid sample into a sealed container for testing. 12.2 Sample processing
12.2.1 Saponification
Weigh 0.25g ~ 10g of prepared sample (nearest to 0.001 g, cholesterol content of about 0.5mg ~ 5mg) in a 250-mL round-bottomed flask. Add 30 mL of
absolute ethanol, 10 mL of 60% potassium hydroxide solution. Well mix. Heat the sample at 100??C magnetic stirring heater for saponification reflux 1h. Shake from time to time so as to prevent sample sticking to the bottle wall. After the saponification ends, use 5 mL of absolute ethanol to rinse its interior from the condenser top. Remove the round-bottomed flask. Cool to room temperature with running water.
12.2.2 Extraction
Quantitatively transfer of all saponified liquid in a 250-mL separatory funnel. Use 30 mL of water to rinse the round-bottomed flask in 2~3 times. Merge the washing liquid into the separatory funnel. Then use 40 mL of petroleum ether - anhydrous ether mixture (1+1, volume ratio) to rinse the round-bottomed flask in 2~3 times. Then merge into the separatory funnel. Shake 2 min, place still and stratify. Transfer the water phase. Combine the three organic phases. Wash the extract to neutral with 100 mL of water each time. Swirl gently for the first time of water washing to prevent emulsification. The extract shall be dehydrated with about 10 g of anhydrous sodium sulfate to a 150-mL flask.
12.2.3 Concentration
Evaporate the extract in the flat-bottomed flask to near dryness under vacuum condition. Use absolute ethanol to dissolve and set volume to 5 mL. The solution passes through a 0.45 ??m filter membrane. Collect the filtrate in the sample bottle. Determine with high performance liquid chromatograph.
The pretreatment of different sample requires blank test at the same time. 12.3 Determination
12.3.1 Instrument reference conditions
a) Chromatographic column. C18 reversed-phase column, column length
17.4.2 Cholesterol standard working solution (100 ??g/mL). pipette 10 mL of cholesterol standard stock solution (1.0 mg/mL); use glacial ac...

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