GB 5413.35-2010 English PDF

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GB 5413.35-2010: National food safety standard -- Determination of β-carotene in foods for infants and young children, milk and milk products
Status: Obsolete

Standard ID	USD	BUY PDF	Lead-Days	Standard Title (Description)	Status
GB 5413.35-2010	319	Add to Cart	3 days	National food safety standard -- Determination of β-carotene in foods for infants and young children, milk and milk products	Obsolete

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Basic data

Standard ID: GB 5413.35-2010 (GB5413.35-2010)
Description (Translated English): National food safety standard -- Determination of ��-carotene in foods for infants and young children, milk and milk products
Sector / Industry: National Standard
Classification of Chinese Standard: C53;X82
Classification of International Standard: 67.100.30
Word Count Estimation: 8,840
Date of Issue: 2010-03-26
Date of Implementation: 2010-06-01
Quoted Standard: GB/T 6682
Regulation (derived from): Circular of the Ministry of Health (2010)7
Issuing agency(ies): Ministry of Health of the People's Republic of China
Summary: This Chinese standard specifies the infant foods and milk products Determination of ��- carotene. This standard applies to infant foods and milk products Determination of ��- carotene.

GB 5413.35-2010: National food safety standard -- Determination of β-carotene in foods for infants and young children, milk and milk products

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National food safety standard.Determination of β-carotene in foods for infants and young children, milk and milk products National Standards of People's Republic of China National Food Safety Standard Determination of infant foods and dairy products β- carotene National food safety standard Determination of β-carotene in foods for infants and young children, milk and milk products People's Republic of China Ministry of Health issued Issued on. 2010-03-26 2010-06-01 implementation

Foreword

This portion of the reference standard of the International Society of analysts (AOAC) 2005.7 β-Carotene in supplement and raw material square law. Appendix A of this standard is a normative appendix, Appendix B is an informative annex. This standard is the first release. National Food Safety Standard Determination of infant foods and dairy products β- carotene

1 Scope

This standard specifies the method for determination of infant foods and dairy products β- carotene. This standard applies to infant food and dairy products Determination of β- carotene.

2 Normative references

The standard file referenced in the application of this standard is essential. For dated references, only the edition date of the note Apply to this standard. For undated references, the latest edition (including any amendments) applies to this standard. Principle 3 After saponification the sample, so that β- carotene becomes completely free state. After extraction with petroleum ether, reversed phase chromatography, external standard.

4 Reagents and materials

Unless otherwise specified, the reagents used in this method are analytically pure water GB/T 6682 provided a water. 4.1 α- amylase. enzyme activity ≥1.5 U/mg. 4.2 anhydrous sodium sulfate (Na2SO4). 4.3 ascorbic acid (C6H8O6). 4.4 petroleum ether. boiling range 30 ℃ ~ 60 ℃. 4.5 methanol (CH4O). chromatography. 4.6 acetonitrile (C2H3N). chromatography. 4.7 chloroform (CHCl3). chromatography. 4.8 n-hexane (C6H14). chromatography. 4.9 Ethanol. volume fraction of 95%. 4.10 potassium hydroxide solution. solid potassium hydroxide 250 g, 200 mL of water was added to dissolve. Pro preparation before use. 4.11 β- carotene standard. 4.12 β- carotene standard solution. 4.12.1 β- carotene standard stock solution (500 μg/mL). Weigh accurately β- carotene standard (4.11) 50 mg (accurate to 0.1 mg), with n-hexane (4.8) and dilute to 100 mL brown volumetric flask. Note. β- carotene standard stock solution requires the following -10 ℃ dark storage, use a period not exceeding three months. Standard stock solution before use to be corrected, specific operations, see Appendix A. 4.12.2 β- carotene intermediate standard solution (100 μg/mL). accurate Pipette 10.0 mL from β- carotene standard stock solution (4.12.1) in Brown solution in 50 mL volumetric flask with n-hexane (4.8) to the mark. 4.12.3 β- carotene standard working solution. β- carotene from the intermediate standard solution (4.12.2) accurately Pipette 0.50,1.00,2.00 respectively, 3.00,4.00 mL solution was transferred to five 100 mL brown volumetric flask with n-hexane (4.8) to the mark, to give a concentration of 0.5, 1.0, 2.0, 3.0, 4.0 μg/mL series of standard working solution. 5. Apparatus 5.1 high performance liquid chromatography with UV detection. 5.2 rotary evaporator. 5.3 temperature magnetic stirrer. 20 ℃ ~ 80 ℃. 5.4 Centrifuge. ≥5000 rpm rev/min. 5.5 Analysis of balance. a sense of the amount of 0.1 mg. 5.6 nitrogen blowing instrument. 5.7 Incubator. 60 ℃ ± 2 ℃. Step 6 Analysis 6.1 Sample Processing 6.1.1 starch sample Weigh mixed sample of about 5 g of solid or liquid sample of about 50 g (average accurate to 0.0001 g) in 250 mL flask was added 1.0 g ascorbic acid (4.3), solid sample required 50 mL45 ℃ ~ 50 ℃ is dissolved in water and mix well. Join lg α- amylase (4.1) to After nitrogen flask covered with cork, set 60 ℃ ± 2 ℃ incubator (5.7) in the enzyme 30 min. 6.1.2 Non-starch sample Weigh mixed sample of about 10 g of solid or liquid sample of about 50 g (average accurate to 0.0001 g), placed in 250 mL Erlenmeyer flask, Was added 1.0 g ascorbic acid (4.3), solid sample of water required to dissolve 50 mL 45 ℃ ~ 50 ℃ and mix well. 6.2 Preparation of test solution 6.2.1 Saponification. A 100 mL 95% ethanol (4.9) was added to the sample solution, mixed well and added to 25 mL of potassium hydroxide solution (4.10) Mix into the magnetic bar, filled with nitrogen discharged air, covered with rubber plug. The 1000 mL beaker was added about 300 mL of water, the beaker On temperature magnetic stirrer (5.3), and when the water temperature controlled at 53 ℃ ± 2 ℃, the flask into a beaker with magnetic stirring for about saponification After 45 min, taken immediately cooled to room temperature. 6.2.2 extract. saponification was fully transferred to 500 mL separatory funnel, was added 100 mL petroleum ether (4.4), shaking gently, exhaust, Stoppered, shaken at room temperature for 10 min Still stratification, the aqueous phase was transferred to another separating funnel as described above for the second extraction. The combined organic phase was washed with water to nearly neutral. The organic phase (4.2) filtration dehydration over anhydrous sodium sulfate. The filtrate income 500 mL flask at Rotary evaporator (5.2) on at 40 ℃ ± 2 ℃ of dry nitrogen conditions to nearly evaporated (never allow evaporated). Dissolve residue with petroleum ether (4.4) Slag and transferred to a 10 mL volumetric flask. Pipette accurately 2.0 mL petroleum ether from the flask in 10 mL screw cap tube, blowing at 40 ℃ ± 2 ℃ nitrogen blowing instrument (5.6) on dry. The test tube was added 1.0 mL of n-hexane (4.8) and shaken to dissolve the residue. The tubes were centrifuged at not less than 5000 rev/min (5.4) 10 min, measured at room temperature to remove standing. NOTE. during experimental operation should pay attention to dark. 6.3 Determination 6.3.1 Reference chromatographic conditions Column. C18 column, 250 mm × 4.6 mm, 5 μm, or equivalent performance of the column. Mobile phase. Chloroform - nitrile - methanol = 31 285, 1 L of the mobile phase was added 0.4 g of ascorbic acid by 0.45 μm membrane filter Reserve. Flow rate. 2.0 mL/min. Detection wavelength. 450 nm. Column temperature. 35 ℃ ± 1 ℃. Injection volume. 20 μL. 6.3.2 Draw the standard curve Respectively β- carotene standard working solution (4.12.3) into the liquid chromatograph (chromatogram see Appendix B), to obtain the peak area. With Peak area of the vertical axis, β- carotene standard working solution concentration as the abscissa standard curve. 6.3.3 Sample Determination The test solution (6.2.2) into the liquid chromatograph to obtain the peak area to obtain test solution β- carotene concentration of the standard curve.

7 expression analysis

Sample β- carotene according to formula (1). ×× = cX (1) Where. X-- sample β- carotene content, in micrograms per hundred grams (μg/100g); c - a standard curve obtained from test solution β- carotene concentration, in micrograms per milliliter (μg/mL); m - mass of the sample in grams (g); 10-- liquid sample volume. Note. This calculation result is smooth, the total amount of trans β- carotene. The arithmetic mean of two under the same condition of independent determination results indicated that the results to three significant figures.

8 Precision

Two independent determination results under the absolute difference in repeatability condition must not exceed 10% of the arithmetic mean.

9 Other

The standard detection limit of 2 μg/100g.

Appendix A

(Normative) Standard concentration correction method β- carotene standard stock solution after preparation needs to be corrected, as follows. Take β- carotene standard stock solution (at a concentration of 500 μg/mL) 10 μL, injection cuvette containing 3.0 mL of hexane, and mix. Measurement Given its absorbance cuvette thickness of 1 cm, hexane is blank, the incident light wavelength of 450 nm, measured parallel to 3 times, and the mean value. Press (2) to calculate the concentration of the solution. × = AX 01.0 01.3 (2) Where. X - β- carotene solution concentration, in micrograms per milliliter (μg/mL); A - β- carotene UV absorbance value; E - β- carotene in hexane solution, the incident light wavelength of 450 nm, cuvette thickness of 1 cm, the solution concentration of 1 mg/L The absorption coefficient was 0.2638; 01.0 01.3 - Determination during the conversion factor of dilution.

Appendix B

(Informative) FIG standard LC B.1 β- carotene standard liquid chromatograph β- carotene standard liquid chromatography is shown in Figure B.1. Figure B.1 β- carotene standard chromatograms ......

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