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GB 5009.83-2016 PDF English

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GB 5009.83-2016: [Including 2025XG1] National food safety standard - Determination of Carotene in Foods
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GB 5009.83: Historical versions

Standard IDUSDBUY PDFDeliveryStandard Title (Description)Status
GB 5009.83-2016120 Add to Cart Auto, 9 seconds. [Including 2025XG1] National food safety standard - Determination of Carotene in Foods Valid
GB/T 5009.83-2003319 Add to Cart 3 days Determination of carotene in foods Obsolete

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GB 5009.86   GB/T 5009.81   GB/T 5009.78   GB 5009.89   

GB 5009.83-2016: [Including 2025XG1] National food safety standard - Determination of Carotene in Foods


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GB NATIONAL STANDARD OF THE PEOPLE’S REPUBLIC OF CHINA National Food Safety Standard - Determination of Carotene in Foods [Including AMD1 / 2025XG1] Issued on. DECEMBER 23, 2016 Implemented on. JUNE 23, 2017 Issued by. National Health and Family Planning Commission of the PRC; China Food and Drug Administration.

Table of Contents

Foreword... 3 1 Scope... 4 2 Principles... 4 3 Reagents and materials... 4 4 Instruments and equipment... 6 5 Analytical procedures... 6 6 Analysis results expression... 11 7 Precision... 13 8 Others... 13 Appendix A Calibration method for concentration of standard solution... 15 Appendix B Confirmation of retention time of β-carotene isomers and calculation of chromatographic purity of all-E-β-carotene... 17 Appendix C Liquid chromatogram of carotene... 19 Appendix D Percentile absorption coefficients of carotene... 21 Amendment No. 1 [2025XG1]... 22

Foreword

This Standard replaces GB 5413.35-2010 “National Food Safety Standard - Determination of β-carotene in foods for infants and young children, milk and milk products”, GB/T 5009.83-2003 “Determination of carotene in foods”, and NY/T 82.15-1988 “Method for determination of fruit juice - Determination of β- carotene”. As compared with GB 5413.35-2010, the main changes of this Standard are as follows. - The standard’s name is changed to “National Food Safety Standard - Determination of Carotene in Foods”; - ADD pretreatment methods for common foods; - ADD the chromatographic conditions where α-carotene and β-carotene need to be differentiated; - MODIFY the results expression of carotene. National Food Safety Standard - Determination of Carotene in Foods

1 Scope

This Standard specifies the method for determination of carotene in foods. This Standard’s chromatographic condition I is applicable to the determination of α-carotene, β-carotene, and total carotene in foods. The chromatographic condition II is applicable to the determination of β-carotene in foods.

2 Principles

The sample is saponified to release the carotene to a free state. After using petroleum ether to extract dichloromethane and dilute, ADOPT reversed phase chromatography to separate and external standard method to quantify.

3 Reagents and materials

Unless otherwise stated, the reagents used in this method are analytically pure. The water is the Grade I water specified in GB/T 6682. 3.1 Reagents 3.1.1 α-amylase. Enzyme activity≥1.5 U/mg. 3.1.2 Papain. Enzyme activity≥5 U/mg. 3.1.3 Potassium hydroxide (KOH). 3.1.9 Chloroform (CHCl3). chromatographically pure. 3.1.10 Methyl tert-butyl ether [CH3OC(CH3)3]. chromatographically pure. 3.1.11 Dichloromethane (CH2Cl2). chromatographically pure. 3.1.12 Anhydrous ethanol (C2H6O). guaranteed reagent. 3.2 Preparation of reagent Potassium hydroxide solution. WEIGH 500 g of solid potassium hydroxide; ADD 500 mL of water to dissolve. Prepared before use. 3.3 Standards 3.4 Preparation of standard solutions 3.4.1 Standard stock solution of α-carotene (500 μg/mL). Accurately WEIGH 50 mg (accurate to 0.1 mg) of α-carotene standard; ADD 0.25 g of BHT; USE dichloromethane to dissolve; TRANSFER to a 100 mL brown volumetric flask and DILUTE to volume. PROTECT it from light at below -20 °C. 3.4.2 Standard intermediate solution of α-carotene (100 μg/mL). Accurately PIPETTE 10.0 mL of standard stock solution of α-carotene into a 50 mL brown volumetric flask; and USE dichloromethane to dilute to volume. 3.4.4 Standard intermediate solution of β-carotene (100 μg/mL). Accurately PIPETTE 10.0 mL of standard stock solution of β-carotene into a 50 mL brown volumetric flask; and USE dichloromethane to dilute to volume. 3.4.5 Mixed standard working solution of α-carotene and β-carotene (for chromatographic condition I). 3.4.6 Standard working solution of β-carotene (for chromatographic condition II). Accurately PIPETTE 0.50 mL, 1.00 mL, 2.00 mL, 3.00 mL, 4.00 mL, and 10.00 mL of standard intermediate solution of β-carotene into 6 100 mL brown volumetric flasks.

4 Instruments and equipment

4.1 Homogenizer. 4.2 High speed pulverizer. 4.3 Constant temperature oscillating water bath. The temperature control precision is ±1 °C. 4.4 Rotary evaporator. 4.7 High performance liquid chromatograph (HPLC). with UV detector.

5 Analytical procedures

Note. Light shall be avoided during the entire test operation. 5.1 Preparation of samples Samples such as grains, beans, and nuts, etc. need to be crushed, ground, and sieved (The hole diameter of sieve plate is 0.3 mm~0.5 mm). 5.2 Treatment for samples 5.2.1 Common food samples 5.2.1.2 Saponification ADD 25 mL of potassium hydroxide solution and CAP the stopper. PLACE in a constant temperature oscillating water bath which has been preheated to 53 °C±2 °C to saponify for 30 min. REMOVE, LET stand, and COOL to room temperature. 5.2.2 Food samples with β-carotene added 5.2.2.2 Saponification TAKE the pretreated sample; ADD 75 mL of anhydrous ethanol, SHAKE well; ADD 25 mL of potassium hydroxide solution; and CAP the bottle stopper. 5.3 Sample extraction TRANSFER the saponification solution to a 500 mL separatory funnel; ADD 100 mL of petroleum ether, gently SHAKE, EXHAUST, and CAP the stopper. After oscillating at room temperature for 10 min, LET it stand for stratification; 5.4 Chromatographic determination 5.4.1 Chromatographic condition I (applicable to the determination of α- carotene, β-carotene, and total carotene in foods) 5.4.1.1 Reference chromatographic conditions Reference chromatographic conditions are listed below. 5.4.1.2 PLOT the standard curve of α-carotene, CALCULATE all-E-β- carotene response factor INJECT the mixed standard working solution of α-carotene and β-carotene into HPLC (for chromatogram, SEE Figure C.1); qualitative based on the retention time; DETERMINE the peak areas of the isomers of α-carotene and β-carotene. For α-carotene, according to the concentration and peak area of the series of standard working solutions, TAKE the concentration as the abscissa, the peak area as the ordinate, to plot standard curve; and CALCULATE regression equation. 5.4.2 Chromatographic condition II (applicable to the determination of β- carotene in foods) 5.4.2.1 Reference chromatographic conditions Reference chromatographic conditions are listed below. 5.4.2.2 Making of standard curve INJECT the standard working solution of β-carotene into HPLC (for chromatogram, SEE Figure C.2); qualitative based on the retention time; DETERMINE the peak area. 5.4.2.3 Sample determination Under the same chromatographic conditions, INJECT the solutions to be determined into liquid chromatograph separately, to perform HPLC analysis; qualitative based on the retention time; USE the external standard method of peak area to quantify.

6 Analysis results expression

6.1 Chromatographic condition I The content of α-carotene in the sample shall be calculated according to the equation (2).

7 Precision

The absolute difference between the two independent determination results, obtained under repeated conditions, shall not exceed 10% of the arithmetic mean.

8 Others

When the sample weighing quantity is 5 g, the detection limit of α-carotene and β-carotene is 0.5 μg/100 g, and the limit of quantitation is1.5 μg/100 g.

Appendix A

Calibration method for concentration of standard solution A.1 Calibration of standard stock solution of α-carotene INJECT 10 μL of standard stock solution of α-carotene (The concentration is approximately 500 μg/mL) into a cuvette containing 3.0 mL of n-hexane, and MIX well. The cuvette has a thickness of 1 cm. TAKE n-hexane as the blank; the incident light has a wavelength of 444 nm; DETERMINE the absorbance value; DETERMINE in parallel for 3 times and TAKE the average.

Appendix B

Confirmation of retention time of β-carotene isomers and calculation of chromatographic purity of all-E-β-carotene Note. When chromatographic condition I is used for the determination of β-carotene, the retention time of β-carotene isomers needs to be determined, and the chromatographic purity of β-carotene standard solution shall be corrected. B.1 Reagent Iodine solution (I2). 0.5 mol/L. B.2 Preparation of reagents B.4 Calculation of chromatographic purity of all-E-β-carotene standard solution ......
Source: Above contents are excerpted from the full-copy PDF -- translated/reviewed by: www.ChineseStandard.net / Wayne Zheng et al.
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