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GB 5009.84-2016

Chinese Standard: 'GB 5009.84-2016'
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Standard ID GB 5009.84-2016 (GB5009.84-2016)
Description (Translated English) (Food safety national standard - Determination of Vitamin B1 in Food)
Sector / Industry National Standard
Classification of Chinese Standard X09
Word Count Estimation 10,129
Date of Issue 2016-08-31
Date of Implementation 2017-03-01
Older Standard (superseded by this standard) GB/T 9695.27-2008; GB/T 7628-2008; GB/T 5009.84-2003; GB 5413.11-2010
Regulation (derived from) Announcement of the State Administration of Public Health and Family Planning 2016 No.11

GB 5009.84-2016
National food safety standard -
Determination of vitamin B1 in foods
食品中维生素 B1的测定
Issued by. National Health and Family Planning Commission of the
People's Republic of China
Table of Contents
Foreword ... 3 
1 Scope ... 4 
2 Principles ... 4 
3 Reagents and materials ... 4 
4 Instruments and equipment ... 6 
5 Analytical procedures ... 6 
6 Expression of analytical results ... 8 
7 Precision... 9 
8 Others ... 9 
9 Principles ... 9 
10 Reagents and materials... 10 
11 Instruments and equipment ... 12 
12 Analytical procedures ... 13 
13 Expression of analytical results ... 15 
14 Precision ... 16 
15 Others ... 16 
Appendix A High performance liquid chromatogram of vitamin B1 standard
derivatives ... 17 
This standard replaces GB/T 5009.84-2003 “Determination of thiamine (vitamin
B1) in foods”, GB 5413.11-2010 “National food safety standard - Determination
of vitamin B1 in foods for infants and young children, milk and milk products”,
GB/T 7628- 2008 “Determination of vitamin B1 in cereals”, GB/T 9695.27-2008
“Meat and meat products - Determination of vitamin B1 content”.
As compared with GB/T 5009.84-2003, the main changes of this standard are
as follows.
- CHANGE the standard name into “National food safety standard -
Determination of Vitamin B1 in foods”;
- ADD the high performance liquid chromatography as the first method, USE
the fluorescence spectrophotometry as the second method;
- MODIFY the expression of detection limit, ADD the quantitative limits of
- ADD the qualitative identification method of chloride ion in the artificial
zeolite pretreatment;
- ADD the color change characteristics of the solution when using the
bromocresol green as the indicator;
- DELETE the Figure 1 (reaction bottle) and Figure 2 (base salt exchange
tube) structure;
- ADD the weighing amount of artificial zeolite when it is expressed in wet
National food safety standard -
Determination of vitamin B1 in foods
1 Scope
This standard specifies the method for determining the vitamin B1 in foods by
the high performance liquid chromatography and fluorescence
This standard applies to the determination of vitamin B1 content in food.
Method 1. High performance liquid chromatography
2 Principles
Samples are hydrolyzed, neutralized, then rehydrated in dilute hydrochloric acid
medium at constant temperature. The hydrolyzate is derivatized by alkaline
potassium ferricyanide solution, extracted with n-butanol, separated by C18
reversed-phase column, detected by high performance liquid chromatography-
fluorescence detector, quantified by external standard method.
3 Reagents and materials
Unless otherwise indicated, the reagents used in this method are of analytical
pure, the water is level I water as specified in GB/T 6682.
3.1 Reagents
3.1.1 n-butanol (CH3CH2CH2CH2OH).
3.1.2 Potassium ferricyanide [K3Fe (CN)6].
3.1.3 Sodium hydroxide (NaOH).
3.1.4 Hydrochloric acid (HCl).
3.1.5 Sodium acetate (CH3COONa • 3H2O).
3.1.6 Glacial acetic acid (CH3COOH).
3.1.7 Methanol (CH3OH). Chromatographic pure
5.1.3 Other solid samples with lower moisture content. for example, the grain
having a moisture content at about 15%, TAKE about 100 g of sample, USE
crusher to crush the sample to obtain the power of constant uniformity, MAKE
determination in time or PRESERVE it in refrigerator.
5.2 Preparation of specimen solution
5.2.1 Test solution extraction
WEIGH 3 g ~ 5 g (accurate to 0.01 g) of solid sample or 10 g ~ 20 g of liquid
sample, PLACE it into a 100 mL conical flask (with a soft stopper), ADD 60 mL
of 0.1 mol/L hydrochloric acid solution, SHAKE it uniformly, PLUG the soft
stopper, PLACE it in the autoclave at 121 °C for 30 min. After finishing
hydrolysis and cooling it to below 40 °C, TAKE it out, SHAKE it gently for several
times; USE the pH indicator, USE 2.0 mol/L sodium acetate solution to adjust
the pH to about 4.0, ADD 2.0 mL (the consumption amount is appropriately
adjusted based on different enzyme activity) of mixed enzyme solution, SHAKE
it uniformly; PLACE it in an incubator at 37 °C for overnight (about 16 h);
TRANSFER all the enzyme solution into a 100 mL volumetric flask, USE water
to make its volume reach to the mark, SHAKE it uniformly, CENTRIFUGE or
FILTER it, TAKE the supernatant to prepare for use.
5.2.2 Test solution derivatization
Accurately PIPETTE 2.0 mL of supernatant or filtrate into a 10 mL test tube,
ADD 1.0 mL of alkaline potassium ferricyanide solution, VORTEX it uniformly,
accurately ADD 2.0 mL of n-butanol, VORTEX it again to mix it uniformly for 5
min, LET it be standing for about 10 min or CENTRIFUGE it; after it is fully
stratified, ABSORB the n-butanol phase (upper layer), USE a 0.45 μm organic
microporous membrane to filter it, TAKE 2 mL of filtrate and PLACE it into a 2
mL brown sample vial to prepare for analysis. If the concentration of vitamin B1
in the test solution exceeds the maximum concentration in the linear range, it
shall take the supernatant to dilute it for several times for re- derivatization, AND
make sample injection again.
TAKE another 2.0 mL of standard series working solution, MAKE it subject to
derivatization synchronously with the test solution.
Note 1. Derivatives are stable within 4 h at room temperature.
Note 2. The operation process of 5.2.1 and 5.2.2 shall be avoided in bright light
Note 3. As for the dried pepper and other samples, when the extract is
determined after direct derivatization, the recovery rate of vitamin B1 is relatively
low. After the extract is purified by the artificial zeolite, the recovery rate of the
c - The concentration of vitamin B1 calculated from the standard curve of the
test solution (extract), in micrograms per milliliter (μg/mL);
V - Test solution (extract) constant volume, in milliliters (mL);
f - Dilution factor of test solution (supernatant) before derivatization;
m - Mass of the specimen, in grams (g).
The calculation results are expressed as the arithmetic mean of two
independent determinations obtained under repeatability conditions, the result
is retained with three significant figures.
Note. The measured thiamine content determined in the specimen is multiplied
by the conversion factor of 1.121, to obtain the content of thiamine
7 Precision
The absolute difference between two independent determinations obtained
under repeatability conditions shall not exceed 10% of the arithmetic mean.
8 Others
When the amount of weighing sample is 10.0 g, in accordance with the constant
volume of this standard method, the detection limit of vitamin B1 in food is 0.03
mg/100 g, the limit of quantitation is 0.10 mg/100 g.
Method 2. Fluorescence spectrophotometry
9 Principles
Thiamin is oxidized to thiopyrimidine in alkaline potassium ferricyanide sol......
Related standard: GB 5009.91-2017    GB 5009.93-2017
Related PDF sample: GB 5009.85-2016    GB 5009.83-2016