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GB 5009.84-2016 PDF English

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GB 5009.84-2016: National food safety standard - Determination of vitamin B1 in foods
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GB 5009.84: Evolution and historical versions

Standard IDContents [version]USDSTEP2[PDF] deliveryName of Chinese StandardStatus
GB 5009.84-2016English75 Add to Cart 0-9 seconds. Auto-delivery National food safety standard - Determination of vitamin B1 in foods Valid
GB/T 5009.84-2003English199 Add to Cart 2 days Determination of thiamine (vitamin B1) in foods Obsolete

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GB 5009.84-2016: National food safety standard - Determination of vitamin B1 in foods

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GB NATIONAL STANDARD OF THE PEOPLE’S REPUBLIC OF CHINA National food safety standard - Determination of vitamin B1 in foods [Including 2023XG1] Issued on. AUGUST 31, 2016 Implemented on. MARCH 01, 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 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 No.1 Modification List [2023XG1]... 18

Foreword

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 method; - 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 weight. 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 spectrophotometry. 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). 3.1.10 Amylase. it shall not contain vitamin B1, enzyme activity ≥ 3700 U/g. 3.2 Reagent preparation 3.2.1 Potassium ferricyanide solution (20 g/L). WEIGH 2 g of potassium ferricyanide, USE water to dissolve it, MAKE its volume reach to 100 mL, SHAKE it uniformly. PREPARE it before use. 3.2.2 Sodium hydroxide solution (100 g/L). WEIGH 25g of sodium hydroxide, 3.2.6 Sodium acetate solution (0.05 mol/L). WEIGH 6.80 g of sodium acetate, ADD 900 mL of water to dissolve it, USE glacial acetic acid to adjust the pH between 4.0 to 5.0, ADD water to make the volume reach to 1000 mL. USE 0.45 μm microporous membrane to filter it before use. 3.2.7 Sodium acetate solution (2.0 mol/L). WEIGH 27.2 g of sodium acetate, USE water to dissolve it, MAKE its volume reach to 100 mL, SHAKE it uniformly. 3.3 Standard substances Vitamin B1 standard substance. thiamine hydrochloride (C12H17ClN4OS • HCl)), CAS. 67-03-8, purity ≥ 99.0%. 3.4 Standard solution preparation

4 Instruments and equipment

4.1 High performance liquid chromatograph with fluorescence detector. 4.3 Centrifuge. speed ≥ 4000 r/min. 4.6 Electric oven or autoclave.

5 Analytical procedures

5.1 Specimen preparation 5.1.1 Liquid or solid powder samples. Immediately after the sample is well- mixed, MAKE determination in time or PRESERVE it in refrigerator. 5.1.2 Fresh fruits, vegetables and meat. 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; U 5.3 Instrument reference conditions 5.3.1 Column. C18 reversed-phase column (particle size 5 μm, 250 mm × 4.6 mm) or equivalent. 5.3.2 Mobile phase. 0.05 mol/L sodium acetate solution - methanol (65 + 35). 5.4 Standard curve production INJECT the standard series working solution derivative into the high performance liquid chromatograph to determine the corresponding peak area of vitamin B1, USE the standard working solution concentration (μg/mL) as the abscissa, the peak area as the vertical axis, to draw the standard curve. 5.5 Determination of the specimen solution In accordance with the chromatographic conditions of clause 5.3, INJECT the specimen derivative solution into a high performance liquid chromatograph to obtain the peak area of vitamin B1, CALCULATE the concentration of vitamin B1 in the solution to be determined in accordance with the standard curve.

6 Expression of analytical results

Sample vitamin B1 (by thiamine) content is calculated in accordance with the formula (1).

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.

9 Principles

Thiamin is oxidized to thiopyrimidine in alkaline potassium ferricyanide solution, under the irradiation of ultraviolet light, the thiopyrimidine fluoresces. Under the given conditions where there is no interference from other fluorescent substance, the intensity of this fluorescence is proportional to the amount of thiopyrimidine amount, i.e.,

10 Reagents and materials

Unless otherwise indicated, the reagents used in this method are of analytical pure, the water is the level II water as specified in GB/T 6682. 10.1 Reagents 10.1.1 Butanol (CH3CH2CH2CH2OH). 10.1.8 Artificial zeolite. 10.1.9 Silver nitrate (AgNO3). 10.1.10 Bromocresol green (C21H14Br4O5S). 10.2 Reagent preparation 10.2.1 0.1 mol/L hydrochloric acid solution. PIPETTE 8.5 mL of hydrochloric acid, USE water to dilute it, MAKE its volume reach to 1000 mL, SHAKE it uniformly. 10.2.2 0.01 mol/L hydrochloric acid solution. MEASURE 50 mL of 0.1 mol/L hydrochloric acid solution, USE water to dilute it, MAKE its volume reach to 500 mL, SHAKE it uniformly. 10.2.8 Potassium ferricyanide solution (10 g/L). WEIGH 1 g of potassium ferricyanide, USE water to dissolve it, MAKE its volume reach to 100 mL, SHAKE it uniformly, PRESERVE it in a brown bottle. 10.2.9 Alkaline potassium ferricyanide solution. PIPETTE 4 mL of 10 g/L potassium ferricyanide solution (10.2.8), USE the 150 g/L sodium hydroxide solution to dilute it to 60 mL, SHAKE it uniformly. PREPARE it before use, USE it in dark. ......
Source: Above contents are excerpted from the full-copy PDF -- translated/reviewed by: www.ChineseStandard.net / Wayne Zheng et al.


      

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