GB 5413.26-2010 English PDF

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GB 5413.26-2010: National food safety standard Determination of taurine in foods for infants and young children, milk and milk products
Status: Obsolete

GB 5413.26: Historical versions

Standard ID	USD	BUY PDF	Lead-Days	Standard Title (Description)	Status
GB 5413.26-2010	399	Add to Cart	3 days	National food safety standard Determination of taurine in foods for infants and young children, milk and milk products	Obsolete
GB/T 5413.26-1997	199	Add to Cart	2 days	Milk powder and formula foods for infant and young children--Determination of taurine content	Obsolete

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Basic data

Standard ID: GB 5413.26-2010 (GB5413.26-2010)
Description (Translated English): National food safety standard Determination of taurine 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: 67.100.10
Word Count Estimation: 10,154
Date of Issue: 2010-03-26
Date of Implementation: 2010-06-01
Older Standard (superseded by this standard): GB/T 5413.26-1997
Adopted Standard: AOAC 997.05, IDT
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 taurine. This standard applies to infant foods and milk products Determination of taurine.

GB 5413.26-2010: National food safety standard Determination of taurine in foods for infants and young children, milk and milk products

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National food safety standard Determination of taurine in foods for infans 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 taurine National food safety standard Determination of taurine 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 standard is equivalent to the second law of international Society of Analysts (AOAC) AOAC 997.05 Taurine in powdered milk and powdered infant formulae. This standard replaces GB/T 5413.26-1997 "Milk powder and infant foods - Determination of taurine." This standard compared with GB/T 5413.26-1997, main changes are as follows. - After the original standard methods OPA High Performance Liquid Chromatography for the first statutory law; - Increased chloride former single-column derivatization HPLC method for the second; - The structure of the original standard was revised. - External standard method using the standard curve; - Increase Appendix A standard liquid chromatogram. Appendix A of this standard is an informative annex. This standard replaces the standards previously issued as follows. --GB 5413-1985, GB/T 5413.26-1997. National Food Safety Standard Determination of infant foods and dairy products taurine

1 Scope

This standard specifies the method for determination of infant foods and dairy taurine. This standard applies to the determination of infant foods and dairy products taurine.

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. After the first column derivatization OPA method Principle 3 Phosphoric acid partial sample was dissolved by ultrasonic oscillation extraction and centrifugation, microfiltration membrane filtration column separated by sodium ions, and O-phthalaldehyde (OPA) derivatization reactions, detection, external standard fluorescence detector.

4 Reagents and materials

Unless otherwise specified, the reagents were of analytical grade water as a water GB/T 6682 regulations. 4.1 Partial phosphoric acid (HPO3). 4.2 trisodium citrate (Na3C6H5O7 · 2H2O). 4.3 Phenol (C6H6O). 4.4 nitric acid (HNO3). 4.5 methanol (CH3OH). chromatography. 4.6 boric acid (H3BO3). 4.7 Potassium hydroxide (KOH). 4.8 PHTHALALDEHYDS (C8H6O2) (OPA). 4.9 2-mercaptoethanol (C2H6OS). 4.10 Polyoxyethylene monolaurate ether (Brij-35). Taurine 4.11 Standard. Purity ≥99%. 4.12 Partial phosphoric acid solution (10 g/L). Weigh 10.0 g metaphosphoric acid (4.1), dissolved in water and dilute to 1000 mL. 4.13 citrate buffer. Weigh 19.6 g trisodium citrate (4.2), add 950 mL of water dissolved in 1 mL of phenol (4.3), with Nitric acid (4.4) adjusted to pH 3.10 ~ 3.25, the 0.45μm filter membrane. 4.14 - column Derivatization solvent (o-phthalaldehyde solution) 4.14.1 potassium borate solution (0.5 mol/L). Weigh 30.9 g of boric acid (4.6), 26.3 g of potassium hydroxide (4.7) dissolved in water and given Volume to 1000 mL. 4.14.2 PHTHALALDEHYDS derivative solution. Weigh 0.60 g PHTHALALDEHYDS (4.8), dissolved in 10 mL of methanol (4.5), add 0.5 mL 2- mercaptoethanol (4.9) and 0.35 g Brij-35 (4.10), with 0.5 mol/L of potassium borate solution (4.14.1) and dilute to 1000 mL, Through 0.45 μm membrane filter. Pro preparation before use. Taurine 4.15 standard solution 4.15.1 taurine standard stock solution (1mg/mL). Weigh accurately 0.1000 g taurine standard (4.11), dissolved in water and constant volume To 100 mL. Stock solution can be stored for 7 days at 4 ℃. 4.15.2 taurine standard working solution. taurine stock standard solution (4.15.1) diluted with water to prepare a series of standard solutions, standard series Concentration. 0,5,10,15,20 μg/mL. Pro preparation before use. 5. Apparatus 5.1 high performance liquid chromatography with fluorescence detector. After 5.2 column reactor. 5.3 fluorescence derived solvent infusion pump. 5.4 ultrasonic oscillator. 5.5 pH meter. accuracy of 0.01. 5.6 Centrifuge. ≥5000 rpm rev/min. 5.7 0.45 μm microporous membrane. 5.8 Balance. a sense of the amount of 1mg, 0.1mg. Step 6 Analysis 6.1 sample Accurately weighed solid sample 1 g ~ 5 g sample, the liquid sample 5 g ~ 20 g (accurate to 0.01g, the sample containing 5 μg of taurine above), Metaphosphoric acid solution plus 30 mL (4.12) was dissolved, shake it well and transferred to 100 mL volumetric flask; an ultrasonic oscillator oscillation 10 min ~ 15 min, removed after cooling to room temperature, add water to the mark; the sample solution at 5000 r/min centrifugation 10 min, the supernatant through 0.45 μm microporous membrane (5.7) filtered and the filtrate then take the middle to prepare for injection. 6.2 Determination 6.2.1 Reference chromatographic conditions Column. Sodium dedicated amino acid analysis column (250 mm × 4.6 mm) or equivalent column. Mobile phase. citrate buffer (4.13). Mobile phase flow rate. 0.30 mL/min. Fluorescence derived solvent flow rate. 0.30 mL/min. Column temperature. 55 ℃. Detection wavelength. excitation wavelength. 338 nm, emission wavelength. 425 nm. Injection volume. 20 μL. 6.2.2 Standard curve drawing Taurine series of standard working solution (4.15.2) after sequentially derived by said measuring machine Recommended chromatographic conditions, record the chromatogram peak area, Chromatogram See Appendix A. The peak area of the vertical axis, the concentration of abscissa, the standard curve. 6.2.3 Determination of test solution The test solution according to the above chromatographic conditions recommended machine determined from the standard curve Richard corresponding concentration test solution.

7 expression analysis

7.1 Calculation Results ×× = VcX (1) Where. X - sample taurine content in mg/100 grams (mg/100g); c - sample concentration of test solution, in micrograms/milliliter (μg/mL); V-- volume sample volume in milliliters (mL); Quality m-- sample in grams (g). 7.2 The results are shown 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. Before the second law chloride single column derivatization Principle 9 Samples dissolved in water, with zinc acetate and potassium ferrocyanide precipitated proteins. Supernatant with dansyl chloride derivatization, derivative through C18 Reversed-phase column separation with UV detector (wavelength 254 nm) or a fluorescence detector (excitation wavelength 330 nm, emission wavelength 530 nm) Detection, external standard. 10 Reagents and materials Unless otherwise specified, the reagents were of analytical grade water as a water GB/T 6682 regulations. 10.1 acetonitrile (CH3CN). chromatography. 10.2 Glacial acetic acid (CH3COOH). 10.3 hydrochloric acid. 10.4 anhydrous sodium carbonate (Na2CO3). 10.5 potassium ferrocyanide [K4Fe (CN) 6]. 10.6 zinc acetate [Zn (CH3COO) 2]. 10.7 sodium [Na (CH3COO)]. 10.8 methylamine hydrochloride (methylamine hydrochloride) (CH3NH2 · HCl). 10.9 dansyl chloride (5-dimethylamino-naphthalene-1-sulfonyl chloride). chromatography. NOTE. dansyl chloride wet sensitive to light and unstable. 10.10 Taurine standard. Purity ≥99%. 10.11 hydrochloric acid (1 mol/L). draw 9 mL of hydrochloric acid (10.3), diluted with water and set the volume to 100 mL. 10.12 precipitant 10.12.1 precipitant Ⅰ. Weigh 15.0 g of potassium ferrocyanide (10.5), dissolved in water and dilute to 100 mL. The precipitating agent at room temperature for 3 Stable for months. 10.12.2 precipitant Ⅱ. Weigh 30.0 g zinc acetate (10.6), dissolved in water and dilute to 100 mL. The precipitating agent at room temperature for 3 Remained stable during the month. 10.13 sodium carbonate buffer (pH 9.5) (80 mmol/L). Weigh 0.424 g of anhydrous sodium carbonate (10.4), add 40 mL of water dissolved, 1 mol/L hydrochloric acid (10.11) adjusted to pH 9.5, water volume to 50 mL. The solution was stable at room temperature for three months. 10.14 dansyl chloride solution (1.5 mg/mL). Weigh 0.15 g dansyl chloride (10.9), acetonitrile (10.1) was dissolved and set the volume to 100mL. Pro preparation before use. 10.15 methylamine hydrochloride solution (20 mg/mL). Weigh 2.0 g methylamine hydrochloride (10.8), dissolved in water and dilute to 100 mL. The solution Stable for three months at 4 ℃. 10.16 sodium acetate buffer (pH4.2) (10 mmol/L). Weigh 0.820 g sodium acetate (10.7), add 800 mL of water was dissolved with ice B. Acid (10.2) adjusted to pH 4.2, water volume to 1000 mL, by 0.45 μm membrane filter. 10.17 taurine standard solution 10.17.1 taurine standard stock solution (1 mg/mL). Weigh 0.1000 g taurine standard (10.10), dissolved in water and dilute to 100 mL. Stock solution can be stored for 7 days at 4 ℃. 10.17.2 taurine working standard solution (ultraviolet detection). taurine standard stock solution (10.17.1) diluted with water to prepare a series of standard Solution, the concentration of the standard series. 0,5,10,15,20 μg/mL. Pro preparation before use. 10.17.3 taurine working standard solution (fluorescence detection). taurine standard stock solution (10.17.2) diluted with water to prepare a series of standard Solution, the concentration of the standard series. 0,0.5,0.10,0.15,0.20 μg/mL. Pro preparation before use. 11 instruments and equipment 11.1 high-performance liquid chromatography with ultraviolet detector or diode array detector or a fluorescence detector. 11.2 pH meter. accuracy of 0.01. 11.3 vortex mixer. 11.4 ultrasonic oscillator. 11.5 Centrifuge. ≥5000 rpm rev/min. 11.6 0.45 μm microporous membrane. 11.7 balance. a sense of the amount of 1mg, 0.1mg. 12 Procedure Sample processing 12.1 12.1.1 Extraction test solution Weigh the solid sample 1 g ~ 5 g, liquid sample 5 g ~ 30 g sample (accurate to 0.01 g, when using a UV detector, a sample containing taurocholic Acid should be in the 1 μg or more, if using fluorescence detector, the sample should contain taurine in 50μg above) in 100 mL volumetric flask, add 80 mL Warm water (50 ℃ ~ 60 ℃) was dissolved, mix well, set the oscillation 10 min ultrasonic oscillators, cooled to room temperature. Add 1.0 mL precipitant Ⅰ (10.12.1), vortex mixing, 1.0 mL precipitant Ⅱ (10.12.2), vortex mixed with water to volume, mix thoroughly, test solution In 5000 r/min centrifugation 10 min, the supernatant spare. The supernatant was stored at 4 ℃ dark for 24 h stable. 12.1.2 derivatized sample solution Draw 1.00 mL above supernatant to 10 mL stoppered glass test tube was added 1.00 mL of sodium carbonate buffer (10.13), 1.00 mL Dansyl chloride solution (10.14), mixed at room temperature in the dark derivatization 2 h (1 h after shaking required 1), A hydrochloric acid was added 0.10 mL Amine (10.15) vortex mixed to terminate the reaction, was allowed to stand in the dark until complete precipitation. Supernatant through 0.45 μm microporous membrane (11.6) Filtered and the filtrate set aside. Derivatives save 48 h at 4 ℃ can be protected from light. Another 1.00 mL standard working solution (10.17.2), synchronized with the test solution is derived. 12.2 Determination 12.2.1 reference chromatographic conditions Column. C18 reverse phase column (particle size 5 μm, 250 mm × 4.6 mm) or equivalent column. Mobile phase. 10 mmol/L sodium acetate buffer (10.16) - acetonitrile (10.1) = 7030. Flow rate. 1.00 mL/min. Column temperature. room temperature. Detection wavelength. UV detector or diode array detector. 254 nm; Or fluorescence detector. excitation wavelength. 330 nm; emission wavelength. 530 nm. Injection volume. 20 μL. 12.2.2 standard curve drawing Taurine series of standard working solution (UV detection) (10.17.2) or taurine series of standard working solution (fluorescence detection) (10.17.3) Derivatives successively by said measuring machine Recommended chromatographic conditions, record the chromatogram peak area chromatogram see Appendix A. The peak area of the vertical axis, The concentration of abscissa, the standard curve. 12.2.3 Determination of test solution The derivatives according to the test solution recommended above chromatographic conditions machine determined from the standard curve Richard corresponding concentration test solution. 13 analysis results presentation Taurine content in the sample according to equation (2). ×× = VcX (2) Where. X-- sample taurine content in mg/100 grams (mg/100g); Sample concentration c-- test solution, in micrograms/milliliter (μg/mL); V-- sample volume by volume, in milliliters (mL); Quality m-- sample in grams (g). The arithmetic mean of two under the same condition of independent determination results indicated that the results to three significant figures. 14 Precision Two independent determination results under the absolute difference in repeatability condition must not exceed 10% of the arithmetic mean. Other 15 The standard quantitative limit. When the sample size was 10.00 g, the first normal 0.5 mg/100g, the second method with UV detection is 5 mg/100g, Fluorescence detection is 0.1 mg/100 g.

Appendix A

(Informative) FIG HPLC standard solution Figure A.1 HPLC standard solution O-phthalaldehyde (OPA) post column derivatization liquid chromatogram see Figure A.1. Derivation chloride former single-column liquid chromatogram (UV detector) see Figure A.2. Derivation chloride former single-column liquid chromatogram (fluorescence detection) see Figure A.3. Figure A.1 o-phthalaldehyde (OPA) post column derivatization liquid chromatogram Figure A.2 single front chloride column derivatization liquid chromatogram (UV detection) nh - 8 .6 AU 0.000 0.010 0.020 0.030 0.040 0.050 0.060 0.070 0.080 Minutes 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 11.00 12.00 13.00 14.00 15.00 ta ur in - 9 .7 EU 0.00 500.00 1000.00 1500.00 2000.00 2500.00 Minutes 0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00 Figure A.3 Derivation chloride former single-column liquid chromatogram (fluorescence detection) .7 EU 0.00 20.00 40.00 60.00 80.00 100.00 120.00 140.00 160.00 Minutes 0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 11.00 12.00 13.00 14.00 15.00 ......

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