GB 28310-2012 English PDF

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GB 28310-2012: Food additive β-carotene (fermentation)
Status: Valid

Standard ID	USD	BUY PDF	Lead-Days	Standard Title (Description)	Status
GB 28310-2012	229	Add to Cart	3 days	Food additive β-carotene (fermentation)	Valid

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Basic data

Standard ID: GB 28310-2012 (GB28310-2012)
Description (Translated English): Food additive ��-carotene (fermentation)
Sector / Industry: National Standard
Classification of Chinese Standard: C54;X40
Classification of International Standard: 67.220.20
Word Count Estimation: 10,180
Regulation (derived from): Ministry of Health Bulletin No. 7 of 2012
Issuing agency(ies): Ministry of Health of the People's Republic of China
Summary: This Chinese standard applies to filamentous fungi trispora (Blakeslea trispora) derived from the fermentation of food additive ��- carotene.

GB 28310-2012: Food additive β-carotene (fermentation)

---This is a DRAFT version for illustration, not a final translation. Full copy of true-PDF in English version (including equations, symbols, images, flow-chart, tables, and figures etc.) will be manually/carefully translated upon your order.
Food additive β-carotene (fermentation) National Standards of People's Republic of China National standards for food safety Food Additives β-carotene (Fermentation) 2012-04-25 release 2012-06-25 Implementation Issued by the Ministry of Health of the People's Republic of China National standards for food safety Food Additives β-carotene (Fermentation)

1 Scope

This standard applies to the filamentous fungus Bacillus thuringiensis (Blakesleatrispora) fermentation of food additives derived from β-carotene. 2 molecular formula, structural formula, relative molecular mass 2.1 Molecular formula C40H56 2.2 Structural formula 2.3 Relative molecular mass 536.88 (according to.2007 International relative atomic mass)

3 technical requirements

3.1 sensory requirements. should be consistent with the provisions of Table 1. Table 1 sensory requirements The project requires a test method Color dark red to brown red State crystalline or crystalline powder Take appropriate sample in a clean, dry white porcelain dish Natural light, observe the color and state 3.2 Physical and chemical indicators. should be consistent with the provisions of Table 2. Table 2 Physical and chemical indicators Item Index Test Method Total beta-carotene content (calculated as C40H56), w /% ≥ 96.0 Appendix A, A.3 Absorbance ratio A455/A483 1.14 ~ 1.19 A455/A340 ≥ 0.75 Appendix A, A.4 Burning residue, w /% ≤ 0.2 Appendix A A.5 Ethanol, w /% ≤ Ethyl acetate, w /% ≤ 0.8 (alone or both) Isopropanol, w /% ≤ 0.1 Isobutyl acetate, w /% ≤ 1.0 Appendix A, A.6 Lead (Pb)/(mg/kg) ≤ 2 GB 5009.12 Total arsenic (in As)/(mg/kg) ≤ 3 GB/T 5009.11 Note. The commercial β-carotene product should be in line with this standard β-carotene as raw material, can be added in line with the requirements of food additives quality requirements Rubber, antioxidant and/or edible vegetable oil, dextrin, starch, the total β-carotene content and absorbance ratio meet the identification value.

Appendix A

Testing method A.1 General provisions The reagents and water used in the standard, when not specified in other requirements, refer to the analytical reagent and the third class specified in GB/T 6682-2008 water. Standard titration solution used in the test, the standard solution for the determination of impurities, preparations and products, without any other requirements, GB/T 601, GB/T 602, GB/T 603. The solution used in the test refers to water when it is not specified with the formulation of the solvent Solution. A.2 Identification test A.2.1 Solubility test A.2.1.1 Reagents and materials A.2.1.1.1 Ethanol. A.2.1.1.2 Vegetable oil. A.2.1.2 Analysis steps At room temperature to take a certain amount of samples were added to a certain amount of water, ethanol and vegetable oil, shake 0.5min ~ 5min, observe the sample Of the dissolution of the situation. β-carotene is almost insoluble in water and ethanol [(solute/solvent) < (1/10000)], slightly soluble in vegetable oil [(1/100) (Solute/solvent) < (1/1000)]. A.2.2 carotenoid test A.2.2.1 Reagents and materials A.2.2.1.1 Acetone. A.2.2.1.2 5% sodium nitrite solution. Weigh 50g sodium nitrite, add water to dissolve, dilute and set to 1000mL. A.2.2.1.3 0.5 mol/L sulfuric acid solution. A.2.2.2 Analysis steps Weighed a certain amount of sample dissolved in acetone, the sample of the acetone solution color in the continuous drop of 5% sodium nitrite solution and 0.5mol/L Sulfuric acid solution gradually disappear. A.3 Determination of total β-carotene content A.3.1 Reagents and materials A.3.1.1 Trichloromethane. A.3.1.2 Cyclohexane. A.3.2 Instruments and equipment UV-visible spectrophotometer. A.3.3 Analysis steps A.3.3.1 Preparation of Solution A. Accurately weighed 0.05g sample, accurate to 0.0001g, placed in 100mL volumetric flask, add 10mL of chloroform solution, with Cyclohexane volume to scale. Take this sample solution 5.0mL in 100mL brown volumetric flask, with cyclohexane volume to the mark, shake, that is, too. A.3.3.2 Solution B Preparation Take solution A5.0mL in 50mL brown volumetric flask, with cyclohexane volume to the mark, shake, that is, too. A.3.3.3 Determination Place the solution B in a 1 cm cuvette, make a blank control with cyclohexane, and use a UV-Vis spectrophotometer at 455 nm ± 2 nm And the absorbance at the maximum absorption wavelength. Absorbance should be controlled between 0.3 to 0.7, or should adjust the concentration of sample solution, and then again Determination of absorbance. A.3.4 Calculation of results The total β-carotene content is expressed in terms of mass fraction w1 of β-carotene (C40H56), expressed in terms of formula (A.1) w1 = c × (A.1) Where. A --- the actual determination of the sample solution absorbance value; c - the value of the test solution concentration in grams per milliliter (g/mL); 2500 - Percentage of β-carotene in cyclohexane (E1 m). The experimental results are based on the arithmetic mean of the parallel measurement results. The absolute difference between the two independent determinations obtained under repeatability conditions The value should not exceed 1.5% of the arithmetic mean. A.4 Determination of absorbance ratio A.4.1 Reagents and materials A.4.1.1 Trichloromethane. A.4.1.2 Cyclohexane. A.4.2 Instruments and equipment UV-visible spectrophotometer. A.4.3 Analysis steps A.4.3.1 Preparation of sample solution Solution A was prepared with A.3.3.1 and Solution B was prepared with A.3.3.2. A.4.3.2 Determination of A455/A483 The solution B was placed in a 1 cm cuvette, and the mixture was taken as a blank control with cyclohexane. At a wavelength of 455 nm and a wavelength of 483 nm, The ratio of luminosity (A), A455/A483 should be between 1.14 and 1.19. A.4.3.3 Determination of A455/A340 The solution B and the solution A were placed in a 1 cm cuvette, and the cyclohexane was used as a blank control, and the solution B was measured at a wavelength of 455 nm (A455), the absorbance (A340) of solution A was measured at a wavelength of 340 nm, and the ratio of A455/A340 should be not less than 0.75. A.5 Determination of burning residue A.5.1 Reagents and materials sulfuric acid. A.5.2 Analysis steps Weigh about 2g sample, accurate to 0.0001g, placed in the 800 ℃ ± 25 ℃ burning to constant weight of the crucible, slowly heated with a small fire To the sample completely carbonized, let cool, add about 1.0mL sulfuric acid to make it moist, low temperature heating to sulfuric acid steam removed, into the high temperature furnace, in the 800 ℃ ± 25 ℃ burning to constant weight. A.5.3 Calculation of results The burning residue is expressed in terms of mass fraction w2 and the value in%, calculated according to formula (A.2) w2 = m1-m2 m x 100% (A.2) Where. m1 - the mass of the residue and the empty crucible, in grams (g); m2 --- the value of the quality of the empty crucible, in grams (g); m --- the value of the sample to be weighed, in grams (g). The experimental results are based on the arithmetic mean of the parallel measurement results, and the absolute difference between the results of the second parallel measurement is not more than 0.05%. A.6 Determination of ethanol, ethyl acetate, isopropanol and isobutyl acetate A.6.1 Reagents and materials A.6.1.1 Ethanol. A.6.1.2 isopropanol. A.6.1.3 Ethyl acetate. A.6.1.4 isobutyl acetate. A.6.1.5 N, N-Dimethylformamide (DMF). Chromatographically pure. A.6.2 Instruments and equipment Gas chromatograph with hydrogen flame ion detector (FID) and headspace sampler. A.6.3 Reference chromatographic conditions A.6.3.1 Column. DB-624 capillary column, 30m x 0.53mm, film thickness 3.0μm; or other equivalent column. A.6.3.2 Carrier gas. helium or nitrogen. A.6.3.3 Carrier gas flow. 4.8 mL/min. A.6.3.4 Inlet temperature. 250 ° C. A.6.3.5 Column temperature. 50 ° C, raised to 60 ° C at 1 ° C/min, then raised to 115 ° C at 9.2 ° C/min and then at 35 ° C/min to 220 ° C Keep for 6min. A.6.3.6 Detector temperature. 270 ° C. A.6.3.7 Split ratio. 5. 1. A.6.3.8 Injection volume. 1 mL dosing ring. A.6.4 Headspace conditions A.6.4.1 Headspace bottle temperature. 100 ° C. A.6.4.2 Dosing ring temperature. 110 ° C. A.6.4.3 Transmission line temperature. 120 ° C. A.6.4.4 Headspace Bottle Balance Time. 50min. A.6.4.5 Gas phase cycle time. 30.0 min. A.6.4.6 Pressing time. 0.2min. A.6.4.7 Dosing ring filling time. 0.2 min. A.6.4.8 Dosing ring equilibration time. 0.05 min. A.6.4.9 Injection time. 1.0 min. A.6.4.10 Top empty bottle pressure. 95.15 kPa (13.8 psi). A.6.5 Analysis steps A.6.5.1 Preparation of control solution A.6.5.1.1 Preparation of the control solution. 34 μL of pure ethanol, 4.5 μL of isopropyl alcohol, 30 μL of ethyl acetate and 38 μL of isobutyl acetate, In a 100 mL volumetric flask, diluted with DMF to the mark, shake, remove 3.0mL to 10mL headspace bottle, cover. The control solution Containing 0.2686 mg/mL ethanol, 0.035325 mg/mL isopropyl alcohol, 0.2703 mg/mL ethyl acetate, isobutyl acetate 0.3325 mg/mL. Note. ethanol density 0.790g/mL, isopropyl alcohol density 0.785g/mL, ethyl acetate density 0.901g/mL, isobutyl acetate density 0.875g/mL. A.6.5.1.2 Sensitivity Solution Preparation. Extract 5.0 mL of the control solution in a 50 mL volumetric flask, dilute to the mark with DMF, shake Uniform, remove 3.0mL to 10mL headspace bottle, cover. The solution contained 0.02686 mg/mL ethanol (corresponding to the concentration in the sample About 0.08%), isopropanol 0.0035325 mg/mL (corresponding to a concentration of about 0.01% in the sample), ethyl acetate 0.02703 mg/mL (Corresponding to a concentration of about 0.08% in the sample) and 0.03325 mg/mL of isobutyl acetate (corresponding to a concentration of about 0.1% in the sample). A.6.5.2 Preparation of sample solution Accurately weigh the sample about 0.1g to 10mL headspace bottle, add 3.0mL DMF dilution, cover, shake, this is the sample solution. A.6.5.3 System Adaptability Requirements Remove the above A.6.5.1 control solution 3.0mL in 6 10mL headspace bottles, respectively, the injection, the number of the theoretical number of peaks should not Less than 5000, the relative standard deviation of the peak area shall not exceed 10.0%, the sensitivity of the main peak signal to noise ratio should not be less than 10. A.6.5.4 Determination Respectively, take the control solution and sample solution headspace injection, with the external standard method to calculate the sample of ethanol, isopropyl alcohol, ethyl acetate and isobutyl acetate content. A.6.6 Calculation of results The contents of ethanol, isopropanol, ethyl acetate and isobutyl acetate are expressed in terms of mass fraction wi, respectively, and the values are expressed in% (A.3). wi = AX x cR AR × cX × 100% (A.3) Where. AX --- the value of the peak area of the solvent measured in the chromatogram of the sample solution; cR --- the value of the corresponding solvent concentration in the control solution in milligrams per milliliter (mg/mL); AR - the value of the peak area of the solvent measured in the chromatogram of the control solution; cX - the concentration of the sample solution in milligrams per milliliter (mg/mL). The experimental results are based on the arithmetic mean of the parallel measurement results. The absolute difference between the two independent determinations obtained under repeatability conditions The value should not exceed 20% of the arithmetic mean. ......

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