|
|
||||||||||||||||||
QB/T 4576-2023 PDF EnglishSearch result: QB/T 4576-2023_English: PDF (QB/T4576-2023)
QB/T4576-2023 (QBT4576-2023): PDF in EnglishQB/T 4576-2023 QB LIGHT INDUSTRY STANDARD OF THE PEOPLE’S REPUBLIC OF CHINA ICS 67.180 CCS X 69 Replacing QB/T 4576-2013 Sodium hyaluronate 透明质酸钠 ISSUED ON. DECEMBER 20, 2023 IMPLEMENTED ON. JULY 01, 2024 Issued by. Ministry of Industry and Information Technology of the PRC Table of Contents Foreword... 3 1 Scope... 5 2 Normative references... 5 3 Terms and definitions... 6 4 Molecular formula, relative molecular mass, structural formula... 6 5 Requirements... 7 6 Test method... 8 7 Inspection rules... 10 8 Marking, packaging, transportation, storage... 11 Appendix A (Normative) Sodium hyaluronate content - Spectrophotometer method 13 Appendix B (Normative) High-performance liquid chromatography method for sodium hyaluronate content... 16 Appendix C (Informative) High-performance liquid chromatogram of sodium hyaluronate... 19 Sodium hyaluronate 1 Scope This document specifies the identification, sensory, physical and chemical, safety and other requirements of sodium hyaluronate; describes the corresponding test methods; specifies the inspection rules, marking, packaging, transportation, storage content; gives the information of molecular formula, relative molecular mass, structural formula. This document applies to the production, inspection, and sale of sodium hyaluronate produced by fermentation of Streptococcus equi subspecies zooepidemicus with glucose, yeast powder, peptone, etc. as raw materials. 2 Normative references The contents of the following documents constitute essential clauses of the text through normative references in the text. Among them, for dated references, only the version corresponding to that date applies to this document; for undated references, the latest version (including all amendments) applies to this document. GB/T 191 Packaging - Pictorial marking for handling of goods GB/T 601 Chemical reagent - Preparations of reference titration solutions GB/T 602 Chemical reagent - Preparations of standard solutions for impurity GB/T 603-2023 Chemical reagent - Preparations of reagent solutions for use in test methods GB 4789.2 National food safety standard - Microbiological examination of food. Aerobic plate count GB 4789.3 National food safety standard - Food microbiological examination - Enumeration of coliforms GB 4789.15 National food safety standard - Food microbiological examination - Enumeration of moulds and yeasts GB 5009.3 National food safety standard - Determination of moisture in food GB 5009.4 National food safety standard - Determination of ash in foods GB 5009.11 National food safety standards - Determination of total arsenic and Determine according to Appendix A or Appendix B, using Appendix A as the arbitration method. 6.5 Moisture Make measurement according to the method 1 "Direct drying method" of GB 5009.3, where the drying time is 4 h. 6.6 Ash Make measurement according to the method 1 "total ash" of GB 5009.4. 6.7 pH 6.7.1 Water Distilled water without carbon dioxide. It is prepared according to 5.1.1.7 of GB/T 603- 2023. 6.7.2 Instruments and equipment pH meter. Accuracy of 0.01. Magnetic stirrer. Thermostatic water bath. Accuracy of ±0.5 °C. 6.7.3 Determination steps Weigh 0.1 g of sodium hyaluronate sample (accurate to 0.01 g). Add 100 mL of distilled water without carbon dioxide. Dissolve by magnetic stirring or heating in a 40 °C water bath. Measure the pH of the solution with a pH meter. The result is rounded to one decimal place. 6.8 Lead (measured as Pb) It is determined according to the method of GB 5009.12. 6.9 Arsenic (measured as As) It is determined according to the method of GB 5009.11. 6.10 Mold and yeast It is determined according to the method of GB 4789.15.If the sample cannot be dissolved, add an appropriate amount of sterile hyaluronidase to help the sample dissolve. 6.11 Total colony count 7.4 Exit-factory inspection The exit-factory inspection items are sensory requirements, sodium hyaluronate content (on a dry basis), pH, moisture, total colony count, mold and yeast, and coliform group. 7.5 Type inspection The type inspection items are all the items specified in the requirements of this document. Generally, type inspection is carried out every six months. Type inspection shall also be carried out in any of the following situations. a) Where there are major changes in raw and auxiliary materials; b) Where key processes or equipment are changed; c) When the new trial products is produced or when the production is restored after production suspension of 3 months; d) Where there is a big difference between the exit-factory inspection and the last type inspection results; e) Where the national market supervision agency needs to conduct random inspections according to relevant requirements. 7.6 Judgment rules If the inspection results show that one item of the product does not meet the requirements of this document, samples shall be taken from twice the amount of packaging for re-inspection; the re-inspection results shall prevail. If there is still one unqualified item, the batch of products shall be judged as unqualified products. If the inspection results show that two or more items of the product do not meet the requirements of this document, the batch of products shall be judged as unqualified products. 8 Marking, packaging, transportation, storage 8.1 Marking The outer packaging marking shall comply with the provisions of GB/T 191.Pre- packaged product labels shall comply with the provisions of GB 7718.For signs with special requirements, they shall be marked as required by the purchaser. 8.2 Packaging The packaging materials that meet the packaging requirements of the corresponding industry products shall be used and can only be used after passing the inspection. Strictly seal to prevent the product from absorbing moisture and leaking. For packaging Appendix A (Normative) Sodium hyaluronate content - Spectrophotometer method A.1 Principle Hyaluronic acid contains N-acetylglucosamine and glucuronic acid in equal molar ratios. Using borax as a catalyst to hydrolyze sodium hyaluronate with sulfuric acid can separate glucuronic acid. Glucuronic acid reacts with carbazole to form an organic complex, which shows a unique purple color; its absorbance is proportional to the mass concentration of glucuronic acid. The content of sodium hyaluronate can be determined by the content of glucuronic acid. A.2 Reagents and solutions A.2.1 Standard. D-glucuronic acid, CAS No.6556-12-3, purity not less than 98%. A.2.2 Concentrated sulfuric acid. High purity. A.2.3 Carbazole. A.2.4 Borax. A.2.5 Anhydrous ethanol. A.2.6 Carbazole ethanol solution (1.25 g/L). Weigh 0.125 g of carbazole (accurate to 0.001 g); dissolve it in 100 mL of anhydrous ethanol; place it in a brown bottle. Store in an explosion-proof refrigerator at 4 °C ~ 8 °C, which has a shelf life of 2 months; or store in the dark at room temperature, which has a shelf life of 15 days. A.2.7 D-glucuronic acid standard solution (0.2 mg/mL). Accurately weigh 20 mg of D- glucuronic acid standard (accurate to 0.0001 g according to actual purity); place it in a 100 mL volumetric flask; add water to dissolve and dilute to the mark; shake well for later use. A.2.8 Borax-sulfuric acid solution (0.025 mol/L). Weigh 4.77 g of borax (accurate to 0.001 g); dissolve it in 500 mL concentrated sulfuric acid; store it in a sealed glass narrow-necked bottle for later use. A.3 Instruments and equipment A.3.1 Vortex mixer. A.3.2 Spectrophotometer. Capable of measuring absorbance at 530 nm. A.4 Drawing of standard curve Measure 0.5 mL, 1.0 mL, 1.5 mL, 2.0 mL, 2.5 mL of glucuronic acid standard solution (A.2.7) respectively; place in a 10 mL volumetric flask; add water to dilute to the scale, to obtain standard solutions with mass concentrations of 10 μg/mL, 20 μg/mL, 30 μg/mL, 40 μg/mL, 50 μg/mL. Take 6 stoppered graduated test tubes; add 1.0 mL of standard solutions of different mass concentrations respectively; then add 5.0 mL of borax sulfuric acid solution respectively; cool in an ice bath for 15 min. Shake gently first; then mix thoroughly with a vortex mixer. Heat the test tube in boiling water for 10 min; remove and cool to room temperature in an ice water bath or running water. Add 0.2 mL of carbazole solution (A.2.6) to the cooled test tube; mix well; heat in a boiling water bath for another 15 min; cool to room temperature. Use a 10 mm cuvette to measure the absorbance at a wavelength of 530 nm; draw a standard curve, using the absorbance as the ordinate and the mass concentration as the abscissa. A.5 Analysis steps A.5.1 Weigh about 0.1 g (m, accurate to 0.0001 g) of sodium hyaluronate sample in a stoppered conical flask; add water to 100 g (w1, accurate to 0.01 g); stir magnetically until fully dissolved. Weigh about 4.0 g (w2, accurate to 0.01 g) of the above solution in a 50 mL (V) volumetric flask; dilute to the mark with water; shake well. A.5.2 Take 1 mL of sample solution; add 5 mL of borax-sulfuric acid solution; cool it in an ice bath for 15 min. Shake gently first; then mix thoroughly with a vortex mixer. Heat the test tube in boiling water for 10 min; then cool to room temperature in an ice water bath or running water. Add 0.2 mL of carbazole test solution; mix well; heat in a boiling water bath for another 15 min; then cool to room temperature. Measure the absorbance at a wavelength of 530 nm, using a spectrophotometer with a 1 cm cuvette. A.6 Calculation The sodium hyaluronate content (on a dry basis) is calculated according to formula (A.1). Wherein. X1 - Sodium hyaluronate content in the sample (on a dry basis), in grams per hundred grams (g/100 g) ρ1 - According to the absorbance of the sample, the corresponding mass concentration of glucuronic acid as found from the standard curve, in micrograms per milliliter (μg/mL); .......Source: https://www.ChineseStandard.net/PDF.aspx/QBT4576-2023 |