Standard No.:	GB 5009.32-2016
English Name:	National Food Safety Standard - Determination of 9 Antioxidants in Food
Chinese Name:	食品安全国家标准 食品中9种抗氧化剂的测定
Professional Classification:	GB    National Standard
Source Content Issued by:	National Health and Family Planning Commission; China Food and Drug Administration
Issued on:	2016-12-23
Implemented on:	2017-6-23
Status:	valid
Superseding:	GB/T 5009.32-2003 Determination of propylgallate in oils and fats GB/T 23373-2009 Determination of butylated hydroxyl anisole(BHA), dibutylated hydroxyl toluene(BHT) and tertiary butyl hydroquinone(TBHQ) in foods
Target Language:	English
File Format:	PDF
Word Count:	8000 words
Translation Price(USD):	240.0
Delivery:	via email in 1 business day

GB 5009.32-2016 National food safety standard - Determination of nine antioxidants in foods 1 Scope This standard specifies five determination methods for nine antioxidants in food: propyl gallate (PG), 2,4,5-trihydroxybutyrophenone (THBP), Tert-butylhydroquinone (TBHQ), Nordihydroguaiaretic acid (NDGA), Butylhydroxyanisole (BHA), 2,6-Di-tert-butyl-4-hydroxymethylphenol (Ionox-100), Octyl gallate (OG), Butylated hydroxytoluene (BHT), and Dodecyl gallate (DG). The methods include high-performance liquid chromatography, liquid chromatography-tandem mass spectrometry, gas chromatography-mass spectrometry, gas chromatography, and colorimetry. The liquid chromatography method in this standard is applicable for the determination of PG, THBP, TBHQ, NDGA, BHA, BHT, Ionox-100, OG, and DG in foods; the liquid chromatography-tandem mass spectrometry method is applicable for the determination of THBP, PG, OG, NDGA, and DG in foods; the gas chromatography-mass spectrometry method is applicable for the determination of BHA, BHT, TBHQ, and Ionox-100 in foods; the gas chromatography method is applicable for the determination of BHA, BHT, and TBHQ in foods; the colorimetric method is applicable for the determination of PG in oils and fats. Method I High-performance liquid chromatography 2 Rationale Oil/fat samples are dissolved in an organic solvent and purified using gel permeation chromatography (GPC); solid food samples are dissolved in n-hexane, extracted with acetonitrile, and purified using a solid-phase extraction column. They are determined by high-performance liquid chromatography and quantified by the external standard method. 3 Reagents and materials Unless otherwise specified, chromatographically pure reagents and Class-I water (defined in GB/T 6682) are adopted for the purpose of this method. 3.1 Reagents 3.1.1 Formic acid (HCOOH). 3.1.2 Acetonitrile (CH3CN). 3.1.3 Methanol (CH3OH). 3.1.4 n-Hexane (C6H14): analytical grade, redistilled. 3.1.5 Ethyl acetate (CH3COOCH2CH3). 3.1.6 Cyclohexane (C6H12). 3.1.7 Sodium chloride (NaCl): analytical grade. 3.1.8 Anhydrous sodium sulfate (Na2SO4): analytical grade, calcined at 650℃ for 4 h, stored in a desiccator, and cooled for further use. 3.2 Preparation of reagents 3.2.1 Acetonitrile-saturated n-hexane solution: Add acetonitrile to n-hexane until saturation. 3.2.2 n-Hexane-saturated acetonitrile solution: Add n-hexane to acetonitrile until saturation. 3.2.3 Eethyl acetate and cyclohexane mixed solution (1+1): Mix 50 mL of ethyl acetate with 50 mL of cyclohexane. 3.2.4 Acetonitrile and methanol mixed solution (2+1): Mix 100 mL of acetonitrile with 50 mL of methanol. 3.2.5 Saturated sodium chloride solution: Add sodium chloride to water until saturation. 3.2.6 Formic acid solution (0.1+99.9): Transfer 0.1 mL of formic acid into a 100 mL volumetric flask and dilute to the scale. 3.3 Standard substance 3.3.1 Butylhydroxyanisole: with a purity ≥ 98%. 3.3.2 Butylated hydroxytoluene: with a purity ≥ 98%. 3.3.3 Octyl gallate: with a purity ≥98%. 3.3.4 Dodecyl gallate: with a purity ≥98%. 3.3.5 propyl gallate: with a purity ≥98%. 3.3.6 Nordihydroguaiaretic acid: with a purity ≥98%. 3.3.7 2,4,5-Trihydroxybutyrophenone: with a purity ≥98%. 3.3.8 Tert-butylhydroquinone: with a purity ≥ 98%. 3.3.9 2,6-Di-tert-butyl-4-hydroxymethylphenol: with a purity ≥ 98%. Note: English names, molecular formulas, and CAS numbers of these substances are provided in Annex A. 3.4 Preparation of standard solution 3.4.1 Mixed stock standard solution of antioxidants: Accurately weigh 0.1 g (to the nearest 0.1 mg) of solid antioxidant standard substance, dissolve it in acetonitrile in a 100 mL amber volumetric flask, dilute to the scale, and prepare a standard mixed stock solution with a concentration of 1000 mg/L. Store in the dark at 0°C to 4°C.

Contents Foreword i 1 Scope 2 Rationale 3 Reagents and materials 4 Apparatus 5 Analytical procedures 6 Calculation (expression) of analytic result 7 Precision 8 Miscellaneous 9 Rationale 10 Reagents and materials 11 Apparatus 12 Analytical procedures 13 Calculation (expression) of analytic result 14 Precision 15 Miscellaneous 16 Rationale 17 Reagents and materials 18 Apparatus 19 Analysis procedure 20 Expression of analysis results 21 Precision 22 Miscellaneous 23 Rationale 24 Reagents and materials 25 Apparatus 26 Analysis procedure 27 Expression of analysis results 28 Precision 29 Miscellaneous 30 Rationale 31 Reagents and materials 32 Apparatus 33 Analysis procedure 34 Expression of analysis results 35 Precision 36 Miscellaneous Annex A English names, molecular formulas and CAS numbers of 9 antioxidants in food Annex B Reference conditions for gel permeation chromatography purification Annex C Standard liquid chromatogram of 9 antioxidants in food Annex D Monitoring ion pairs, collision energy, residence time and retention time of antioxidants in foods Annex E LC-MS-MS diagram for multiple reaction monitoring of 5 antioxidants (THBP, PG, OG, NDGA, and DG) in foods Annex F Retention time, quantitative ions, qualitative ions, abundance ratio and residence time of antioxidants in foods Annex G GC-MS diagram for selected ion monitoring of four antioxidants in food Annex H Gas chromatograms of standard solutions of three antioxidants (BHA, BHT and TBHQ) in food