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GB 5009.258-2016 (GB5009.258-2016)

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GB 5009.258-2016: PDF in English

GB 5009.258-2016
NATIONAL STANDARD OF THE
PEOPLE’S REPUBLIC OF CHINA
National Food Safety Standard –
Determination of Raffinose in Foodstuffs
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
1 Scope ... 3 
2 Principle ... 3 
3 Reagents and Materials ... 3 
4 Instruments and Equipment ... 4 
5 Analytical Procedures ... 5 
6 Expression of Analytical Result ... 7 
7 Precision ... 7 
8 Others ... 7 
Appendix A Typical Ion Chromatogram of Standard Solution and Sample
Solution ... 8 
National Food Safety Standard -
Determination of Raffinose in Foodstuffs
1 Scope
This Standard specifies the ion chromatography for the determination of raffinose in
foodstuffs.
This Standard is applicable to the determination of raffinose content in infant formula
foods, infant cereal supplementary food, infant canned supplementary food, beverage
and soy flour food.
2 Principle
Adopt the mode of acetonitrile precipitation and high-speed centrifugation to remove
protein and fat in the sample. After solid phase extraction column purification, inject
the sample into ion chromatograph. Take sodium hydroxide solution as the eluent.
Separate through anion exchange column; detect through pulse amperometric
detector. Conduct qualitative determination through the retention time; conduct
quantitative determination through the external standard method.
3 Reagents and Materials
Unless it is otherwise stipulated, all reagents used in this method shall be excellent-
grade purity. Water shall be Grade-1 water stipulated in GB/T 6682.
3.1 Reagents
3.1.1 Acetonitrile (C2H3N): chromatographic purity.
3.1.2 50% sodium hydroxide solution (NaOH): chromatographic purity.
3.1.3 -amylase: enzyme activity  1.5 U/mg.
3.1.4 Nitrogen (N2): purity  99.99%.
3.2 Preparation of Reagents
Sodium hydroxide solution (250 mmol/L): add 500 mL of distilled water, which is filtered
through 0.22 μm nylon filter, to 2 L plastic reagent bottle. Weigh-take 13.1 mL of sodium
hydroxide solution (3.1.2) to below the water surface. Add distilled water to 1,000 mL.
After injecting nitrogen protection, evenly shake it; reserve for later usage.
5 Analytical Procedures
5.1 Sample Preparation
5.1.1 Sample pre-treatment
5.1.1.1 Milk powder, rice flour and soy flour samples: take around 100 g of sample
(other solid samples shall be smashed and grinded first), then, fill it into a container
with lid, which can hold twice the volume of the sample. Through repeatedly shaking
and reversion of the container, the sample could be thoroughly mixed, till the sample
becomes homogeneous.
5.1.1.2 Liquid samples: take around 100 mL of sample, then, fill it into a container with
lid, which can hold twice the volume of the sample. Through repeatedly shaking and
reversion of the container, the sample could be thoroughly mixed, till the sample
becomes homogeneous.
5.1.1.3 Muddy flesh and other semi-solid samples: weigh-take 50.0 g of sample; weigh-
take an equivalent mass of water, then, mix them up. Start vortex mixing, then,
ultrasound for 20 min.
5.2 Sample Treatment
5.2.1 Starch-containing samples, for example, rice flour: weigh-take 1 g ~ 5 g (accurate
to 0.1 mg, contain around 20 mg of raffinose) of thoroughly-mixed sample; place it in
a centrifuge tube. Add 1 g of -amylase. Solid sample needs 50 mL of 45 °C ~ 50 °C
water to be dissolved. After thoroughly mixing it, fill in nitrogen; put on the tube cap.
Place it into a 60 °C ± 2 °C incubator, then, start enzymatic hydrolysis for 30 min. After
cooling it down to room temperature, use water to reach the constant volume of 100
mL.
5.2.2 Other samples, such as milk powder, soy flour, beverage and muddy flesh:
weigh-take 1 g ~ 5 g (accurate to 0.1 mg, contain around 20 mg of raffinose) of
thoroughly-mixed sample; place it in a 100 mL beaker. Add water to dissolve it. Solid
sample needs 50 mL of 45 °C ~ 50 °C water to be dissolved. Transfer it into a 100 mL
volumetric flask, then, use water to dilute to a constant volume.
5.3 Purification
5.3.1 Absorb 5 mL of previously-treated sample solution, then, place it into a 50 mL
conical flask with a plug. Add 10 mL of water; mix it up. Start ultrasound for 30 min.
Then, transfer it into a 50 mL volumetric flask. Add 30 mL of acetonitrile; use water to
dilute to a constant volume. Transfer it into a 50 mL centrifuge tube. At 10,000 r/min,
start centrifugation for 10 min. Take the supernatant for later usage (in accordance with
the content of raffinose in the sample solution, the dilution factor may be adjusted).
......
(Above excerpt was released on 2019-06-18, modified on 2021-06-07, translated/reviewed by: Wayne Zheng et al.)
Source: https://www.chinesestandard.net/PDF.aspx/GB5009.258-2016