GB 15570-2010National food safety standards -- Food additives -- Folic acid (English PDF)

Foreword

This standard replaces GB 15570-1995 "Food Additives folic acid." this standard compared with GB 15570-1995, the main changes are as follows.

- Indicators of folic acid by the mass fraction of 95% to 102% revised to 96% to 102%;

- Residue on ignition <=0.1% by the index revised to <=0.2%;

- Determination of folic acid canceled colorimetric spectroscopy, quantitative methods HPLC internal standard method was modified by the external standard method;

- Improve the moisture determination method.

Appendix A of this standard is a normative appendix, Appendix B is an informative annex.

This standard replaces the standards previously issued as follows.

--GB 15570-1995.

National Food Safety Standard Folic acid food additives

1 Scope

This standard applies to chemical synthesis of folic acid food additives.

2 Normative references

The standard file referenced in the application of this standard is essential. For dated references, only the date of the note Stage version applies to this standard. For undated references, the latest edition (including any amendments) applies this standard.

3 Chemical name, molecular formula, molecular mass and structural formula

4 Technical Requirements

4.1 Sensory requirements. comply with Table 1.

Table 1 Sensory requirements

Project requires test methods Yellow or orange color take appropriate sample is placed in a clean, dry white porcelain dish, natural Light, observe its color and texture. Organizational status crystalline powder

4.2 Physical indicators. to comply with Table 2.

Table 2. Physical and chemical indicators

Item Index Test Method Folic acid (C19H19N7O6, dry basis), w /% 96.0 ~ 102.0 Appendix A A.4 Water, w /% <= 8.5 A.5 in Appendix A Residue on ignition, w /% <= 0.2 A.6 in Appendix A Heavy metals (Pb)/(mg/kg) <= 10 Appendix A A.7 Arsenic (As)/(mg/kg) <= 3 GB/T 5009.76 Appendix A (Informative) Testing method A.1 Safety Tips Reagents The standard test methods used for toxic or corrosive, according to the relevant provisions of the operation, the operator would like to be careful Shen. If splashed on the skin should immediately wash with water, severe cases should be treated immediately. When using a volatile acid, to enter in the hood Row.

A.2 General Provisions this standard reagents and water, did not indicate when the other requirements, refer to analytically pure reagents and GB/T 6682-2008 in Regulation Given three water; test the standard titration solution and other required solution, when it is not specified, according to GB/T 601, GB/T 602, GB/T 603 provisions formulated.

A.3 Identification Test A.3.1 Reagents and materials Sodium hydroxide solution. c (NaOH) = 0.1 mol/L.

A.3.2 Instruments and Equipment UV spectrophotometer, with a 1 cm absorption cell.

A.3.3 UV absorption test Weigh laboratory sample, add sodium hydroxide solution produced per 1 mL containing 10 µg laboratory sample solution by UV spectrophotometry Meter measured at 256 nm ± 2 nm, 283 nm ± 2 nm, 365 nm ± 4 nm wavelength of maximum absorption at 256 nm and 365 Nm wavelength absorbance ratio should be 2.8 to 3.0.

A.4 Determination of folic acid A.4.1 Reagents and materials A.4.1.1 Methanol. HPLC grade.

A.4.1.2 potassium dihydrogen phosphate.

A.4.1.3 potassium hydroxide solution. c (KOH) = 0.1 mol/L.

A.4.1.4 ammonia solution. 0.5 -> 100.

A.4.1.5 folic acid reference substance.

A.4.2 Instruments and Equipment High performance liquid chromatography.

A.4.3 Chromatography conditions Recommended chromatographic columns and typical operating conditions are shown in Table A.1, folic acid typical HPLC diagram in Appendix B, other You can achieve the same degree of separation and efficiency of the column and chromatographic conditions can be used.

Table A.1 column chromatography and typical operating conditions

Column length 25cm, column diameter 4.6mm, octadecyl silane bonded silica gel column.

The mobile phase with 70 mL of potassium dihydrogen phosphate 6.8g potassium hydroxide solution, diluted with water to 850 mL, and the pH adjusted to 6.3 ± 0.1, add 80 mL of methanol, diluted with water to 1000 mL of solution.

A flow rate of 1mL/min.

Detection wavelength 254 nm.

The column temperature was 30 °C, control accuracy ± 1 °C.

A.4.4 Analysis step

A.4.4.1 reference solution.

Weigh about 10 mg of folic acid reference substance, to the nearest 0.02 mg, set 50 mL volumetric flask, add 30 mL aqueous ammonia solution to dissolve After diluted with water to the mark.

A.4.4.2 laboratory sample solution.

Weigh about 10 mg laboratory samples, accurate to 0.02 mg, set 50 mL volumetric flask, add 30 mL aqueous ammonia solution to dissolve After diluted with water to the mark.

A.4.4.3 system suitability test Number of theoretical plates folic acid peak calculation should not be less than 2000, folic acid peak and the adjacent impurity peak separation should be greater than 1.5.

A.4.4.4 Determination

Table A.1 column chromatography and typical operating conditions, the precise amount of 10 µL laboratory sample solution into the liquid chromatograph, record

Record the chromatograms; the other to take folic acid reference solution, the same method.

A.4.5 Calculation Results Folic acid (C19H19N7O6 to count, dry basis) of the mass fraction in 1w and its value is expressed in%, according to equation (A.1) count Operator.

twenty three

100%

A m ww

A mw

Where. m - injection volume of laboratory sample values, expressed in milligrams (mg);

1m - injection volume of folic acid reference value, expressed in milligrams (mg);

2w - Folic acid reference substance (in terms C19H19N7O6, wet basis) of the mass fraction of the value, expressed in%;

3w - Experimental determination of moisture content of products sample mass fraction press A.5 value, expressed in%;

1A - Numerical laboratory sample solution folic acid peak area;

2A - folic acid reference solution peak area values.

Take two parallel determination results of the arithmetic average of the measurement results, the relative deviation of two parallel determination results is not more than 2.0%.

A.5 Determination of Moisture A.5.1 Reagents and materials A.5.1.1 Fischer's test solution.

A.5.1.2 anhydrous methanol.

A.5.2 Instruments and Equipment Fischer's moisture meter.

A.5.3 Analysis step

Press the "People's Republic of China Pharmacopoeia" 2005 edition two Appendix M VIII determination of moisture content determination A first method. It weighs about 0.1 g Laboratory samples, accurate to 0.0002 g, solvent is anhydrous methanol directly measured with a moisture meter, or dry laboratory sample set the dry glass bottle with stopper, add 25mL ~ 30mL of anhydrous methanol continuously shaken (or stirred) under the test solution with Fischer's titration to dissolve Liquid from light yellow to reddish brown, or indicate the end with a dead-stop titration; otherwise blank test.

A.5.4 Calculation Results Laboratory sample moisture content in 3w and its value is expressed in%, according to equation (A.2) Calculated.

() 100% VV Fw

Where. F - Fischer's test solution 1mL per equivalent value of water quality, in milligrams per milliliter (mg/mL);

1V - Numerical laboratory sample consumption Fischer's test solution volume in milliliters (mL);

0V - Numerical blank test solution consumed by Fischer's volume in milliliters (mL);

m - mass of the laboratory sample values in milligrams (mg).

Take two parallel determination results of the arithmetic average of the measurement results, the relative deviation of the two parallel test should be less than 0.2%.

A.6 Determination of residue on ignition A.6.1 Reagents and materials sulfuric acid.

A.6.2 Analysis step

Weigh about 1.0 g laboratory samples, accurate to 0.0002 g, is set to a constant weight has been burning crucible, slowly burning to complete Carbonization, cooled to room temperature; adding sulfuric acid 0.5mL ~ 1mL or moist, heated to a low temperature after the sulfuric acid vapor divisible at 550 °C ± 50 °C Burning completely gray, moved to a dryer, cooled to room temperature, and precisely weighed after set, and then calcined to constant weight at 550 °C ± 50 °C.

A.6.3 Calculation Results the mass fraction of residue on ignition in 4w and its value is expressed in%, according to equation (A.3) Calculated.

twenty three

() 100% m mw

Where. 2m - numerical constant weight of the crucible and residue mass in grams (g);

3m - numerical constant weight crucible mass in grams (g);

m - mass of the laboratory sample values in grams (g).

Take two parallel determination results of the arithmetic average of the measurement results, the two parallel determination results is not more than the absolute difference 0.02%.

A.7 Determination of Heavy Metals A.7.1 Reagents and materials A.7.1.1 nitrate.

A.7.1.2 sulfuric acid.

A.7.1.3 hydrochloric acid.

A.7.1.4 glycerol.

A.7.1.5 ammonium acetate.

A.7.1.6 lead nitrate.

A.7.1.7 thioacetamide.

A.7.1.8 ammonia solution. 400 -> 1000.

A.7.1.9 sodium hydroxide solution. c (NaOH) = 1 mol/L.

A.7.1.10 hydrochloric acid solution. c (HCl) = 2 mol/L.

A.7.1.11 hydrochloric acid solution. c (HCl) = 7 mol/L.

A.7.1.12 ammonia solution. c (NH3 · H2O) = 5 mol/L.

A.7.1.13 phenolphthalein indicator solution. 10g/L ethanol solution.

A.7.1.14 acetate buffer (pH3.5).

Weigh approximately 25 g of ammonium acetate, adding 25 mL dissolved in water, add 7 mol/L hydrochloric acid solution 38 mL, with 2 mol/L hydrochloric acid solution Exact solution or aqueous ammonia solution to adjust the pH to 3.5 (pH meter), diluted with water to 100 mL, that is, too.

A.7.1.15 thioacetamide test solution.

It weighs about 4 g thioacetamide, to the nearest 0.01 g, add water to dissolve into a 100 mL, home refrigerator. Immediately prior to taking 5.0 mL mixture (by a 1 mol/L 15 mL of sodium hydroxide solution, 5.0 mL of water and 20 mL glycerol), plus the aforementioned 1.0 mL Thioacetamide solution was heated on a water bath 20s, cooling, use immediately.

A.7.1.16 lead standard solution.

Weigh about 0.160 g of lead nitrate, accurate to 0.000 2g, placed in 1000 mL volumetric flask, add 5 mL of nitric acid and 50 mL After dissolved in water, dilute to the mark, shake, as the stock solution. Before use, Pipette 10mL ± 0.02mL stock solution, put 100 mL volumetric flask, diluted with water to the mark, shake, that was (per 1mL equivalent to 10 µg of Pb). Configuration and Storage Glassware used shall not contain lead.

A.7.2 Analysis step

Press the "People's Republic of China Pharmacopoeia" 2005 edition Appendix VIII H second test for determination of heavy metals, specific methods as follows.

Take the residue left under A.6, add 0.5 mL of nitric acid, evaporated to dryness, then to nitric oxide vapor is divisible (or take the laboratory sample 1g, Slowly burn to completely carbonized, cooled to room temperature, 0.5 mL ~ 1.0 mL of sulfuric acid, so just moist, with a low temperature heating to sulfuric acid After divisible, add 0.5 mL of nitric acid, evaporated to nitric oxide and vapor divisible, cooled to room temperature, burning at 500 ~ 600 °C to End Full ashing), cooled to room temperature, add 2 mL of hydrochloric acid, add 15 mL water was evaporated to dryness on a water bath, dropping ammonia solution to phenolphthalein means Shown VIS neutral, plus 2 mL acetate buffer (pH3.5), after heat gently to dissolve, displacing A Nessler colorimetric tube, add water to dilute Release into 25 mL; Another laboratory sample preparation reagent solution, evaporated to dryness rear porcelain dish, add 2 mL acetate buffer (pH 3.5) After dissolution with 15 mL water, slightly hot, displacing Nessler colorimetric tube B, add 1mL ± 0.01mL lead standard solution, and then diluted with water Into 25 mL; and then were added to each of 2 mL of thioacetamide test solution B in the two, shake 2min, on the same set of paper, Top-down perspective, color comparison tube B A tube display and no deeper.

Appendix B

(Informative)

Determination of folic acid a typical HPLC chromatogram

Figure B.1 Determination of folic acid in a typical high-performance liquid chromatogram

Folic acid peak