Food additive vitamin B6 testing

1 Safety Reminder

Some of the reagents used in this experimental method are toxic or corrosive, and should be handled according to relevant regulations with caution. If splashed onto the skin, rinse immediately with water, and in severe cases, seek immediate treatment. When using volatile acids, it should be done in a fume hood

2 General regulations,

Unless otherwise specified, only reagents confirmed as analytical pure and Grade III water specified in GB/T6682-2008 shall be used in the analysis.

The standard titration solution, impurity determination standard solution, formulation, and product used in the test method shall be prepared in accordance with the provisions of GB/T601, GB/T602, and GB/T603 unless otherwise specified

3 Identification test

3.1 Reagents and materials

3.1.1  Sodium acetate solution: 200g/L

3.1.2; Boric acid solution: 40g/L

3.1.3 Chloroimino-2,6-dichloroquinone solution: 5g/L ethanol solution

3.1.4; Manganese dioxide

3.1.5; Sulfuric acid

3.1.6 Starch potassium iodide test paper

3.2 Analysis Steps

3.2.1 Color Reaction

Take about 10mg of laboratory sample, dissolve it in 100mL of water, and take 1mL of each sample. Place it in two test tubes, A and B, respectively, with 2 mL of sodium acetate solution added. Add 1mL of water to tube A and 1mL of boric acid solution to tube B. Mix well, and quickly add 1mL of chloroimino-2,6-dichloroquinone solution to tube A. The blue color appears in tube A, but disappears after a few minutes and turns red; Blue color does not appear in tube B

3.2.2 Identification by infrared absorption spectrum

Using potassium bromide compression method, the infrared spectrum of the laboratory sample should be consistent with the control spectrum.

3.2.3 Chloride Identification

Take approximately 0.5g of laboratory sample and place it in a test tube. Add 0.5g of manganese dioxide, mix well, wet with sulfuric acid, and slowly heat to produce chlorine gas. Starch potassium iodide test paper that can be wetted with water will turn blue.

4 Determination of Vitamin B6

4.1 Method Summary

Using crystal violet as an indicator, titrate the sample with a perchloric acid standard solution, and calculate the vitamin B6 content as C8H11NO3 · HCl based on the amount of perchloric acid standard titration solution used.

4.2 Reagents and Materials

4.2.1 Glacial acetic acid

4.2.2 Mercury acetate solution: Take 5g of mercury acetate, grind it fine, and dissolve it into 100mL with warm glacial acetic acid

4.2.3 Crystal violet indicator solution: 5g/L glacial acetic acid

4.2.4 Perchlorate standard titration solution: c (HClO4)=0.1mol/L

4.3 Analysis Steps

Weigh 0.15g of the laboratory sample to the nearest 0.0002g, add 20mL of glacial acetic acid and 5mL of mercury acetate solution, dissolve at warm temperature, cool to room temperature, add 1 drop of crystal violet indicator solution, titrate with a perchloric acid standard titration solution until the solution turns blue-green, and correct the titration results with a blank test

4.4 Result Calculation

The mass fraction w1 of vitamin B6 (calculated as C8H11NO3 ∙ HCl) is expressed in%, and calculated according to formula (1):

In the formula:

V - the volume of perchloric acid standard titration solution (4.2.4) consumed by the test solution, in milliliters (mL)

V0- Volume value of perchloric acid standard titration solution (4.2.4) consumed in the blank test, in milliliters (mL)

C - The accurate value of the concentration of perchloric acid standard titration solution, in moles per liter (mol/L)

m - The numerical value of the sample mass (calculated as dry product), in grams (g)

M - The numerical value of the molar mass of vitamin B6 (C8H11NO3 · HCl), in grams per mole (g/mol) (M=205.64)

Take the arithmetic mean of the two parallel measurement results as the measurement result, and the absolute difference between the two parallel measurement results shall not exceed 0.5%

5 Determination of drying loss

5.1 Analysis steps

Weigh 1.0g of laboratory sample, accurate to 0.0002g, and measure according to GB/T6284 method

5.2 Result Calculation

The mass fraction w2 of the drying reduction is expressed in%, and calculated according to equation (2):

Where:

m2- The total mass of the sample and weighing bottle before drying, in grams (g)

m1- The total mass of the dried sample and weighing bottle, in grams (g)

M - mass of the sample, in grams (g)

Take the arithmetic mean of the parallel measurement results as the measurement result, and the absolute difference between the two parallel measurement results shall not exceed 0.05%

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