GB 5009.86-2016 PDF English
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| Standard ID | Contents [version] | USD | STEP2 | [PDF] delivery | Name of Chinese Standard | Status |
| GB 5009.86-2016 | English | 115 |
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National food safety standard - Determination of Ascorbic Acid in Foods
| Valid |
| GB/T 5009.86-2003 | English | 239 |
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Determination of total ascorbic acid in fruits, vegetables and derived products -- Fluorometric method and colorimetric method
| Obsolete |
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GB 5009.86-2016: National food safety standard - Determination of Ascorbic Acid in Foods---This is an excerpt. Full copy of true-PDF in English version (including equations, symbols, images, flow-chart, tables, and figures etc.), auto-downloaded/delivered in 9 seconds, can be purchased online: https://www.ChineseStandard.net/PDF.aspx/GB5009.86-2016
GB
NATIONAL STANDARD OF THE
PEOPLE’S REPUBLIC OF CHINA
National Food Safety Standard -
Determination of Ascorbic Acid in Foods
Issued on. AUGUST 31, 2016
Implemented on. MARCH 1, 2017
Issued by. National Health and Family Planning Commission of the
People's Republic of China
Table of Contents
Foreword... 3
1 Scope... 4
2 Terms and definitions... 4
3 Principle... 5
4 Reagents and materials... 5
5 Instruments and equipment... 6
6 Analysis steps... 7
7 Expression of analysis results... 8
8 Precision... 9
9 Other... 9
10 Principle... 10
11 Reagents and materials... 10
12 Instruments and equipment... 11
13 Analysis steps... 12
14 Result calculation... 13
15 Precision... 14
16 Other... 14
17 Principle... 14
18 Reagents and materials... 14
19 Determination... 16
20 Result calculation... 16
21 Precision... 17
Annex A L(+)- ascorbic acid, D(+)- ascorbic acid standard chromatogram... 18
Foreword
This Standard replaces GB/T 5009.86-2003 Determination of total ascorbic acid
in fruits, vegetables and derived products - Fluorometric method, GB/T
5009.159-2003 Determination of reductive-form ascorbic acid in foods and GB
6195-1986 Determination of vitamin C in vegetables and fruits (2,6-dechloro-
indophenol titration method).
Compared with GB/T 5009.86-2003, the main changes are as follows.
- modified the standard name to “National Food Safety Standard-
Determination of Ascorbic Acid in Foods”;
- expanded the application scope of the method;
- added the high-performance liquid chromatography and related
quantitation limitations;
- deleted 2,4-dinitrophenylhydrazine method;
- added 2,6-dichloro-indophenol titration;
- modified the structure of the previous edition according to GB/T 20001.4-
2015.
National Food Safety Standard -
Determination of Ascorbic Acid in Foods
1 Scope
This Standard specifies the high-performance liquid chromatography,
fluorescence method, 2,6-dichloro-indophenol titration method for the
determination of ascorbic acid in food.
Method One of this Standard applies to the determination of total amount of
L(+)- ascorbic acid, D(+)- ascorbic acid and L(+)- ascorbic acid in milk powder,
cereals, vegetables, fruits and their products, meat products, vitamin
supplements, jellies, gummies, rice pudding, wine. Method Two is applicable to
the determination of L(+)- total amount of ascorbic acid in milk powder,
vegetables, fruits and their products. Method Three applies to the determination
of L(+)- total amount of ascorbic acid in fruits, vegetables and their products.
2 Terms and definitions
2.1 Ascorbic acid. an organic compound with antioxidant properties. It is also
known as "Vitamin C," one of the essential nutrients in human body.
2.2 L(+)- ascorbic acid. left-right rotation ascorbic acid that has a strong
reduction. It has biological activity on the human body.
2.5 L(+)- total amount of ascorbic acid. total amount of ascorbic acid obtained
when L(+)- dehydroascorbate in the sample is reduced to L(+)- ascorbic acid
OR L(+)- dehydroascorbic acid is oxidized by L(+)- ascorbic acid in the sample.
Method One -- High performance liquid
chromatography
3 Principle
After ascorbic acid in the sample is dissolved with metaphosphoric acid and
extracted by sonication, use ion-pair reagent as mobile phase. Separate it
through reverse phase column. L(+)- ascorbic acid and D(+)- ascorbic acid are
measured directly with a liquid chromatograph (wavelength of 245 nm)
equipped with a UV detector.
4 Reagents and materials
Unless otherwise specified, the reagents used in this method are of analytical
grade pure, water is of grade one specified in GB/T 6682.
4.1 Reagents
4.1.1 Metaphosphoric acid (HPO3)n. content (calculated as HPO3) ≥38%
4.1.7 Methanol (CH3OH). chromatographically pure
4.2 Reagent preparation
4.2.1 Methanophosphoric acid solution (200 g/L). weigh 200 g (to the nearest
of 0.1 g) of metaphosphoric acid (4.1.1), dissolve in water and dilute to 1 L; this
solution can be stored in a 4°C environment for one month.
4.2.4 L-cysteine solution (40 g/L). weigh 4g of L-cysteine; dissolve in water
and dilute to 100 ml. Prepare it when using.
4.3 Standard product
4.3.1 L(+)- ascorbic acid standard product (C6H8O6). purity ≥99%
4.4 Preparation of standard solution
4.4.1 L(+)- ascorbate standard stock solution (1.000 mg/mL). accurately
weigh 0.01 g of L(+)- ascorbic acid standard product (to the nearest of 0.01 mg);
use 20 g/L meta-phosphoric acid solution to set volume to 10 mL.
4.4.2 D(+)- ascorbate standard stock solution (1.000 mg/mL). accurately
weigh 0.01 g of D(+)- ascorbate standard product (to the nearest of 0.01 mg);
use 20 g/L metaphosphoric acid solution to set volume to 10 mL.
5 Instruments and equipment
5.1 Liquid chromatograph. equipped with a diode array detector or UV detector
5.2 pH meter. precision of 0.01
5.5 Centrifuge. speed ≥4000 r/min
5.6 Homogenizer
5.7 Filter membrane. 0.45 μm aqueous film
6 Analysis steps
The entire testing process is under the conditions as dark as possible.
6.1 Sample preparation
6.1.1 Liquid or solid powder sample. after well mixing, it shall be immediately
used for testing.
6.2 Preparation of sample solution
Weigh about 0.5g ~ 2g (to the nearest of 0.001 g) of uniformly mixed solid
sample or homogenized sample with respect to the sample OR pipette 2mL ~
10Ml of liquid sample [making that the sample contains about 0.03mg ~ 6mg of
L(+)- ascorbic acid] into 50mL beaker. Transfer the sample to a 50-mL
volumetric flask with 20 g/L metaphosphoric acid solution (4.2.2).
6.3 Sample solution reduction
Accurately pipette 20 mL of supernatant that is through centrifugation in 50-mL
centrifuge tube. Add into 10 mL of 40 g/L L-cysteine solution (4.2.4).
6.4 Instrument reference conditions
6.4.1 Chromatographic column. C18 column, column length of 250 mm, inner
diameter of 4.6 mm, particle size of 5 μm, or chromatographic column of same
performance.
6.4.4 Flow rate. 0.7 mL/min
6.4.5 Detection wavelength. 245 nm
6.5 Making of standard curve
Respectively determine the ascorbic acid mixed standard series of working
solutions.
6.6 Determination of sample solution
Determine the sample solution. Obtain the concentration (μg/mL) of L(+)-
ascorbate [or D(+)- ascorbate] according to the standard curve.
7 Expression of analysis results
The amount of L(+)- ascorbic acid [or D(+)- ascorbic acid] and the total amount
of L(+)- ascorbic acid in the sample are expressed in milligrams per hundred
grams, calculated according to equation (1).
8 Precision
The absolute difference between two independent determination results
obtained under repeatability conditions shall not exceed 10% of the arithmetic
mean.
9 Other
When the amount of solid sample is 2 g, the detection limits of L(+)- ascorbic
acid and D(+)- ascorbic acid are both 0.5 mg/100g, the limits of quantification
are 2.0 mg/100g.
10 Principle
After the oxidation of L(+)- ascorbic acid to L(+)- dehydroascorbic acid by
activated carbon, it reacts with o-phenylenediamine (OPDA) to generate
fluorescent quinoxaline.
......
Source: Above contents are excerpted from the full-copy PDF -- translated/reviewed by: www.ChineseStandard.net / Wayne Zheng et al.
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