GB/T 22246-2025Determination of pantothenic acid in health foods (English PDF)

5.1 Reagents

5.1.1 Water, Grade 1 water as specified in GB/T 6682.

5.1.2 Hydrochloric acid (HCl).

5.1.3 Sodium hydroxide (NaOH).

5.1.4 Phosphoric acid (H3PO4). 100 mL volumetric flasks, add water to dilute to the scale and mix them well to obtain pantothenic acid series standard working solutions with concentrations of 1 g/mL, 2 g/mL, 4 g/mL, 8 g/mL, 16 g/mL and 32 g/mL. Prepare immediately prior to use.

5.4 Materials Filter membrane. 0.45 m, aqueous.

6.2 Balance. with a division value of

0.001 g and

0.1 mg.

6.3 Constant-temperature oscillating water bath. with an oscillating frequency of 100 r/min 20 r/min.

6.4 Ultrasonic cleaner.

6.5 pH meter. with an accuracy of 0.01.

6.6 High-speed centrifuge. speed not less than 8,000 r/min.

7.1 Specimen Preparation

7.1.1 Solid specimens (hard capsules, tablets and powders, etc.). take at least 20 capsules or at least 5 g of sample (for hard capsules, remove the shells and take the contents; for tablets, crush them), finely grind (if necessary), mix well and set aside.

7.1.2 Solid specimens (jelly candies, etc.). take at least 10 capsules (for mixed specimens of different colors or flavors, evenly conduct the sampling by color or flavor) or at least 10 g of sample, cut into small pieces and set aside.

7.1.3 Semi-solid specimens (soft capsules, etc.). take at least 20 capsules or at least 5 g of sample, remove the shells, take the contents, mix well and set aside.

7.1.4 Liquid specimens (oral liquids, etc.). take 5 minimum-size packages or at least 50 mL of sample (if the sample contains carbonic acid, use ultrasound to remove carbon dioxide), mix well and set aside.

7.2 Specimen Preparation Weigh-take 1 g ~ 3 g of specimen (accurate to

0.001 g), or transfer-take 1 mL ~ 3 mL of specimen, or a volume that obtains approximately 1 mg of pantothenic acid in the specimen. Place it in a 100 mL conical flask. Add 30 mL of warm water at 40 °C ~ 50 °C; vortex or stir it, until the sample is thoroughly dissolved. Perform ultrasound for 20 minutes, and cool to room temperature. Use

0.1 mol/L hydrochloric acid solution (5.1.8),

1.0 mol/L hydrochloric acid solution (5.1.9),

0.01 mol/L sodium hydroxide solution (5.1.10) or

0.1 mol/L sodium hydroxide solution (5.1.11) to adjust the pH to 5.0.Add 5 mL of

0.5 mol/L zinc sulfate solution (5.1.12) and mix it thoroughly. Transfer to a 50 mL volumetric flask, use water to dilute to the scale, mix it thoroughly, and transfer to a centrifuge tube. At 8,000 r/min, centrifuge for 2 min. Pass the supernatant through a 0.45 m filter membrane (5.4) and prepare the filtrate for determination on the machine. NOTE. if necessary, based on the content of the components in the specimen, appropriately increase the dilution factor f to ensure that the mass concentration of pantothenic acid in the specimen is within the range of 1.0 g/mL ~ 32.0 g/mL.

7.3 Chromatographic Reference Conditions The chromatographic reference conditions are as follows:

a) Chromatographic column. C18 (particle size 5 m, 250 mm

4.6 mm), or equivalent chromatographic column;

b) Mobile phase. potassium dihydrogen phosphate solutionacetonitrile (5.1.14);

c) Flow rate. 1 mL/min;

d) Column temperature. 35 °C;

e) Injection volume. 10 L or 20 L;

f) Detection wavelength. 200 nm.