GB 5009.280-2020
Determination of 4-hexylresorcinol residues in national food safety standards (English Version)

Standard No.

GB 5009.280-2020

Language

Chinese, Available in English version

Release Date

2020

Published By

National Health Commission of the People's Republic of China

Latest

GB 5009.280-2020

Scope

This standard specifies the determination of 4-hexyl m-quinol residues in crabs and crabs. This standard is applicable to the determination of 4-hexyl m-quinol residues in crabs. Deer tide samples are extracted with ethyl acetate, B And saturated n-hexane degreasing, using high performance liquid chromatography and fluorescence detector detection, external standard method 3 Reagents and materials Unless otherwise specified, the reagents used in this method are of analytical grade, and the water is the first-grade water specified in GB/T6682 . 3.1 Reagents 3.1.1 Byouyan (CC, HsN): chromatographically pure. 3.1.2 "Methanol (CH; OH): chromatographically pure. 3.1.3 Ethyl acetate (C, HsO,): chromatographically pure. 3.1.4 n-hexane (CCesHi): chromatographically pure. 3.1.5 "Anhydrous sulfuric acid Sodium (NasSO): put anhydrous sodium sulfate in a muffle furnace and burn at 400°C for 4 hours, then put it in a desiccator for later use. 3.2 "Reagent preparation 3.2.1 Ethanol and saturated n-hexane solution: add a certain amount of ethanediol to n-hexane, shake and mix well, after the mixture is clearly separated, the upper layer is ethylene glycol saturated n-hexane. The upper layer 3.2.2 "Methanol-methanol-water mixed bath solution (1+2+1, volume ratio): Measure 100 mL methanol, 200 mL methanol and 100 mL water respectively, mix in a glass beaker Mix well and set aside. 3.3 Standard substance 4-hexyl m-quinol standard substance (Cs HisO,, CAS number: 136-77-6): purity 399%%, or a standard substance that has been certified by the state and granted a standard substance certificate. 3.4 Preparation of standard solution 3.4.1 4-hexyl m-catechol standard stock solution (100 pg/mL): Accurately weigh 10 mg (accurate to 0.01 mg) of 4-hexyl m-techol standard product, and dissolve it with methanol And dilute to 100 mL, mix spoon. Transfer the solution to a glass container and store it at -18°C, valid for 6 months. 1 GB5009.280-2020 3.4.2 14-Hexyl m-catechol standard intermediate solution (10 pg/mL): pipette 1.00 mL of standard stock solution (3.4.1) into a 10 mL volumetric flask, add methanol to volume. Ready to use. 3.4.3 4-hexyl m-quinol standard series working solution: draw 0.02 mL, 0.05 mL, 0.10 mL, 0.50 mL, 1.00 mL, . , Add methanol-ethanol-water mixed solution to make up to the mark. The mass concentrations of the 4-hexyl m-quinol standard series working solutions were 0.020 0 png/mL, 0.050 0 ng/mL, 0.100 ng/mL, 0.500 pg/mL, 1.00 pg/mL, 2.00 pg/mL, respectively. Ready to use. 4 Instruments and Equipment 4.1 Liquid Chromatograph: Equipped with prized light detector. 4.2 "Balance: Sensitivity is 0.01 g and 0.01 mg. 4.3 Rotary hair dryer. 4.4 Vortex mixer. 4.5 Ultrasonic generation required. 4.6" Centrifuge: The speed can reach 4 000 r/min. 4.7 Tissue Shredder. 4.8 Homogenizer. 4.9 Concentrate bottle. 4.10 Organic phase microporous membrane: 0.45 μm. 5 Analysis steps and 5.1 Sample preparation Take about 500 g of the edible part of the representative sample, grind and mix it with a tissue grinder, put it in a clean container as a sample, seal it and make a good mark. Samples were stored at -18. 5.2 Sample extraction Weigh 2 g (accurate to 0.01 g) of sample into a 50 mL centrifuge tube, add 20 mL of ethyl acetate and 3 g of anhydrous sodium sulfate, homogenize for 30 s, centrifuge at 4000 rpm for 3 min, and put the above All the supernatant was transferred to a 100 mL concentrating bottle, and 20 mL of ethyl acetate was added to the residue, homogenized for 30 s, centrifuged at 4 000 r/min for 3 min, and the supernatant was mixed with the supernatant obtained for the first time. Combined in the same concentrated bottle, in a 35°C water bath to hair to near king. 5.3 Sample purification Add 2 mL of methanol-ethanol-water mixed solution to the residue to redissolve, sonicate for 10 s, and vortex for 30 s. , transferred to a 15 mL centrifuge tube, centrifuged at 4 000 rpm for 3 min, discarded the n-hexane coal layer, added 2 mL of ethyl and saturated n-hexane solution to the centrifuge tube again, vortexed for 30 s, centrifuged at 4000 rpm for 3 min, and discarded the upper layer For the n-hexane layer, the lower layer was filtered through a 0.45 pm organic microporous membrane, and the filtrate was put on the machine. 5.4 The blank test shall be carried out according to the measurement steps in 5.2 and 5.3, except that no sample is added. GB5009.280-2020 5.5 Determination 5.5.1 Instrument reference conditions a) Chromatographic column: Cs column, 250 mmxX4.6 mm (id.), particle size 5 pm bp) Mobile phase: B and decahydrate 265 1035 c) Flow rate: 1.0 mL/min; d) Detection wavelength: Excitation wavelength 280 nm, emission flow length 310 nm e) Column temperature: 35 ” CC, uniform” Injection volume: 10 AL. 5.5.2 Preparation of the standard curve: Inject the standard series of working solutions into the high-performance liquid chromatograph respectively, measure the corresponding peak area, take the mass concentration of 4-hexyl m-quinol in the standard series of working solutions as the abscissa, and take the mass concentration of The peak area of sunbiol is the ordinate, and the standard curve is drawn. Please refer to Appendix A for the chromatogram of this standard solution of 4-hexyl m-quinone. 5.5.3 Determination of the sample solution The sample solution was injected into the high performance liquid chromatograph, and the peak area of the analyte was obtained through the retention time qualitatively. According to the standard curve, the mass concentration of 4-hexyl m-catechol in the test solution was obtained. 6 Calculation and expression of results The content of 4-hexyl methanediol in the sample is calculated according to the formula (1): "c-co) Jiu Liu XX1000 Quan 7 X Shang 000 1 In the formula: X" - 4- in the sample The content of hexyl m-catechol, the unit is milligrams per dry gram (mg/kg); c "-the mass concentration of 4-hexyl m-catechol in the sample solution obtained from the standard curve, the unit is micrograms per milliliter ( ug/mL),; cu—the mass concentration of 4-hexyl m-quinol in the blank test obtained from the standard curve, in micrograms per milliliter (ug/mL); V”—the reconstitution volume of the test sample, The unit is milliliter (mL); 1 000 is the unit conversion factor; 77I is the sample weight of the sample, and the unit is gram (g). The calculation result retains 3 significant figures. The two independent results obtained under repeatability conditions The absolute difference of the measurement results shall not exceed 10 parts of the arithmetic mean. 8 Others When the sample weight of shrimp and crab is 2.00 g, the detection limit is 0.0200 mg/kg, and the quantification limit is 0.0500 mg/kg. GB5009. 280-2020 Appendix A4-Hexyl m-catechol standard solution and spiked chromatogram A.1 and the liquid chromatogram of 4-hexyl m-catechol standard solution are shown in Figure A.1. counts 1600 0003 1400 000]1200 000] 1000 000]800 000]600 000]400 000]200 000]

GB 5009.280-2020 history

• 2020 GB 5009.280-2020 Determination of 4-hexylresorcinol residues in national food safety standards

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