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GB 5009.128-2016National food safety standard - Determination of cholesterol in foods (English PDF)

食品安全国家标准食品中胆固醇的测定

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Issued by

National Health and Family Planning Commission of the People's Republic of China

Level / Type

National · Mandatory

Issue date

December 23, 2016

Implementation date

June 23, 2017

Scope

GB 5009.128-2016 is the English-translated version of 食品安全国家标准食品中胆固醇的测定.

GB 5009.128-2016 — 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

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Document preview — GB 5009.128-2016

National Standard of the People's Republic of China

Issued by: National Health and Family Planning Commission of the People's Republic of China

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

3.2 Regent preparation

3.2.1 60% potassium hydroxide solution. weigh 60 g of potassium hydroxide;

slowly add water to dissolve and set volume to 100 mL.

3.2.2 Petroleum ether - anhydrous ether mixture (1+1, volume ratio).

uniformly mix petroleum ether and anhydrous ether of equal volume.

3.3 Standard product

Cholesterol standard (C27H46O, CAS number. 57-88-5). purity >= 99%.

3.4 Preparation of standard solution

4 Instruments and equipment

4.1 Gas chromatograph. equipped with flame ionization detector (FID)

4.4 Saponification device

5 Analysis steps

5.1 Sample preparation

5.1.1 Meat and meat products and other types of solid samples

Take 200 g of edible part of sample to homogenize

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.

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.

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

5.3.2 Production of standard curve

Inject the cholesterol standard series of working solution into the gas

chromatograph. Under the above chromatographic conditions, measure the

response value of the standard solution (peak area). Using concentration as

abscissa, the peak area as ordinate, make the standard curve.

6 Expression of analysis result

The content of cholesterol in the sample is calculated according to equation (1).

7 Precision

The absolute difference between two independent determinations obtained

under repeatability conditions shall not exceed 10% of the arithmetic mean.

8 Other

When the sample volume is 0.5 g, the set volume is 5.0 mL, the detection limit

shall be 0.3 mg/100g, the limit of quantification shall be 1.0 mg/100g.

9 Principle

Saponify the sample by absolute ethanol - potassium hydroxide solution.

Extract by petroleum ether and anhydrous ether mixture. Concentrate the

extract to dry.

10 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.

10.1 Reagents

10.2 Regent preparation

10.2.1 60% potassium hydroxide solution. weigh 60 g of potassium hydroxide;

slowly add water to dissolve and set volume to 100 mL.

10.2.2 Petroleum ether - anhydrous ether mixture (1+1, volume ratio).

uniformly mix petroleum ether and anhydrous ether of equal volume.

10.3 Standard product

Cholesterol standard (C27H46O, CAS number. 57-88-5). purity >= 99%.

10.4 Preparation of standard solution

11 Instruments and equipment

11.1 Homogenizer

11.2 High performance liquid chromatography. equipped with UV detector

or equivalent detector

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This preview omits tables, figures, formulas and parts of the technical clauses. The complete document — 16 pages — is available in the English PDF.

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