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GB/T 37545-2019: Determination of 38 kinds of permitted colorants in cosmetics - High performance liquid chromatography Delivery: 9 seconds. True-PDF full-copy in English & invoice will be downloaded + auto-delivered via email. See step-by-step procedure Status: Valid
Similar standardsGB/T 37545-2019: Determination of 38 kinds of permitted colorants in cosmetics - High performance liquid chromatography---This is an excerpt. Full copy of true-PDF in English version (including equations, symbols, images, flow-chart, tables, and figures etc.), auto-downloaded/delivered in 9 seconds, can be purchased online: https://www.ChineseStandard.net/PDF.aspx/GBT37545-2019 NATIONAL STANDARD OF THE PEOPLE’S REPUBLIC OF CHINA ICS 71.100.70 Y 42 GB 37545-2019 Determination of 38 kinds of permitted colorants in cosmetics - High performance liquid chromatography ISSUED ON: JUNE 04, 2019 IMPLEMENTED ON: JANUARY 01, 2020 Issued by: State Administration for Market Regulation; Standardization Administration of the People’s Republic of China. Table of ContentsForeword ... 3 Introduction ... 4 1 Scope ... 5 2 Normative references ... 5 3 Principle ... 5 4 Reagents and materials ... 5 5 Instruments and equipment ... 7 6 Sample preparation ... 7 7 Analysis steps ... 8 8 Result calculation ... 12 9 Recovery rate and precision ... 13 10 Tolerance ... 13 Appendix A (Informative) Chinese name, detection concentration, detection limit, minimum quantitation concentration and quantitation-limit of 38 permitted colorants ... 14 Appendix B (Informative) Information sheet for 38 permitted colorants ... 16 Appendix C (Informative) Liquid chromatographic separation chromatogram of the standard substances of 38 permitted colorants ... 23 Determination of 38 kinds of permitted colorants in cosmetics - High performance liquid chromatography1 ScopeThis Standard specifies the principle, reagents and materials, instruments and equipment, sample preparation, analysis steps, result calculation, recovery rate and precision, tolerance and other contents for the high performance liquid chromatography determination of 38 kinds of permitted colorants in cosmetics. The high performance liquid chromatography specified in this Standard applies to the quantitative determination of 38 permitted colorants (excluding lakes) in lipsticks, nail polishes, powders and other cosmetics. Refer to Table A.1 in Appendix A for the detection-limit and quantitation-limit of the target of this standard method.2 Normative referencesThe following referenced documents are indispensable for the application of this document. For dated references, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies. GB/T 6682, Water for analytical laboratory use - Specification and test methods3 PrincipleUse the tetrahydrofuran-methanol-water system to simultaneously extract the water- soluble and oil-soluble colorants in cosmetics. After centrifuging to remove the base material, use methanol to redissolve and dilute the extract which has been concentrated by nitrogen blowing; use the reversed-phase high performance liquid chromatographic separation, and the standard curve external standard method for quantitation. For cosmetic samples which are confirmed by high performance liquid chromatography to be out of range or out of limit in using permitted colorants, mass spectrometry confirmation shall be taken when necessary.4 Reagents and materialsUnless otherwise stated, the water used is the grade-1 water specified in GB/T 6682. 4.1 Standard substance: the purity is greater than 90%. See Table B.1 in Appendix B for the Chinese names, common name of the colorant index, CI number, CAS number, molecular formula, structural formula, and relative molecular mass of 38 kinds of permitted colorants. 4.2 Methanol: analytical reagent. 4.3 Methanol: chromatographic pure. 4.4 Formic acid: chromatographic pure. 4.5 Tetrahydrofuran: analytical reagent. 4.6 Dimethyl sulfoxide: analytical reagent. 4.7 Acetonitrile: chromatographic pure. 4.8 Ammonium acetate: analytical reagent. 4.9 Sodium ascorbate: analytical reagent. 4.10 Standard solution diluent: Accurately weigh 0.1 g of sodium ascorbate (4.9); add 5 mL of water to dissolve; continue to add 5 mL of tetrahydrofuran; use methanol (4.3) to fix volume to 100 mL. (Formulate when needed) 4.11 0.5 mol/L ammonium acetate aqueous solution (containing 0.5% sodium ascorbate): Accurately weigh 3.85 g of ammonium acetate (4.8) and 0.5 g of sodium ascorbate (4.9) into a volumetric flask; dissolve in water and dilute to 100 mL. 4.12 0.03 mol/L ammonium acetate aqueous solution (containing 0.075% formic acid): Accurately weigh 2.31 g of ammonium acetate (4.8); first, add water to dissolve; then, continue to add 750 μL of formic acid (4.4) that is accurately pipetted; finally, add water to fix volume to 1 L; shake and mix well for later use. 4.13 Standard stock solution A: Accurately weigh 5 mg (accurate to 0.01 mg) of each standard substance (Food Red 10, Acid Violet 43, Acid Orange 10, Acid Blue 62, Acid Yellow 1, Acid Blue 7, Acid Yellow 11, Food Red 9, Food Blue 1, Food Red 7, Food Yellow 3, Food Red 17, Food Blue 2, Food Green 3 and Food Blue 5, 15 kinds in total) in the brown volumetric flasks, respectively; dissolve in water and fix volume to 10 mL, to prepare into standard stock solutions with a mass concentration of 500 μg/mL; store in the dark at 4 °C ~ 6 °C; the storage period is 3 months. 4.14 Standard stock solution B: Accurately weigh 5 mg (accurate to 0.01 mg) of each standard substance (Acid Red 88, Bromocresol Green, Basic Violet 2, Acid Orange 7, Acid Red 50, Acid Violet 9, Acid Green 9, Acid Black 1, Acid Red 52. Acid Blue 1, Acid Yellow 121, Disperse Yellow 16, Basic Blue 26, Acid Red 98, Solvent Yellow 29, Acid Violet 50, Acid Green 22, 17 kinds in total) in the brown volumetric flasks, respectively; dissolve in methanol (4.3) and dilute to 10 mL, to prepare into standard stock solutions with a mass concentration of 500 μg/mL; store in the dark at 4 °C ~ 6 °C; the storage period is 3 months. 4.15 Standard stock solution C: Accurately weigh 5 mg (accurate to 0.01 mg) of each standard substance (Solvent Red 3, Solvent Yellow 33 and Solvent Green 3, 3 kinds in total) in the brown volumetric flasks, respectively; dissolve in tetrahydrofuran and fix volume to 10 mL, to prepare into standard stock solutions with a mass concentration of 500 μg/mL; store in the dark at 4 °C ~ 6 °C; the storage period is 3 months. 4.16 Standard stock solution D: Accurately weigh 5 mg (accurate to 0.01 mg) of the standard substance (Pigment Red 4, 1 kind in total) in a brown volumetric flask; dissolve in tetrahydrofuran and dilute to 10 mL, to prepare into a standard stock solution with a mass concentration of 500 μg/mL; store in the dark at 4 °C ~ 6 °C; the storage period is 3 months. 4.17 Standard stock solution E: Accurately weigh 5 mg (accurate to 0.01 mg) of each standard substance (Pigment Red 49 and Pigment Red 64, 2 kinds in total) in the brown volumetric flasks, respectively; dissolve in dimethyl sulfoxide and dilute to 10 mL, to prepare into standard stock solutions with a mass concentration of 500 μg/mL; store in the dark at 4 °C ~ 6 °C; the storage period is 3 months. 4.18 0.45 μm microporous polytetrafluoroethylene (PTFE) filter membrane needle filter.5 Instruments and equipment5.1 High performance liquid chromatography - diode array detector. 5.2 High performance liquid chromatography tandem quadrupole mass spectrometer, equipped with electrospray ionization source (ESI) (used for qualitative confirmation). 5.3 Analytical balance: sensitivity of 0.001 g, 0.000 01 g. 5.4 Vortex mixer. 5.5 Ultrasonic cleaner: the power is not less than 300 W. 5.6 Centrifuge: the rotation speed is not less than 5 000 r/min, and the capacity of the centrifugal test tube is 10 mL. 5.7 Pipette or pipettor.6 Sample preparationAccurately weigh 0.2 g (accurate to 0.001 g) of the sample into a 10 mL centrifuge tube with stopper; first, add 6 mL of tetrahydrofuran (4.5); vortex and sonicate for 10 min to disperse the sample and extract the target (if the lipstick sample cannot be dispersed, range or out of limit in using permitted colorants, mass spectrometry confirmation shall be taken when necessary. 7.5 Quantitative determination Pipette the series standard working solutions (7.3); carry out the high performance liquid chromatography according to the chromatographic conditions (7.1); take the mass concentration of the series standard working solutions as the abscissa and the peak area as the ordinate to make a standard curve. Acid Orange 10, Acid Green 22 and Acid Violet 9 (3 kinds) are quantitatively calculated according to the UV absorption at 254 nm; Acid Yellow 1, Acid Yellow 11, Pigment Red 49, Solvent Yellow 33, Acid Yellow 121, Solvent Red 3, Disperse Yellow 16, Solvent Yellow 29 (8 kinds) are quantitatively calculated according to the UV absorption at 416 nm; Food Yellow 3, Acid Orange 7, Acid Violet 43, and Pigment Red 4 (4 kinds) are quantitatively calculated according to the UV absorption at 484 nm; Food Red 9, Food Red 7, Food Red 10, Food Red 17, Acid Red 50, Acid Red 52, Acid Red 98, Acid Red 88, Bromocresol Green, Basic Violet 2, Pigment Red 64 (11 kinds) are quantitatively calculated according to the UV absorption at 514 nm; Acid Black 1, Acid Violet 50 (2 kinds) are quantitatively calculated according to the ultraviolet absorption at 590 nm; Food Blue 1, Food Green 3, Food Blue 2, Acid Blue 1, Food Blue 5, Acid Green 9, Acid Blue 62, Acid Blue 7, Solvent Green 3, Basic Blue 26 (10 kinds) are quantitatively calculated according to the UV absorption at 620 nm. In order to ensure the accuracy of quantitative test, the test process should be completed within 10 h.8 Result calculationThe content of the target in the sample solution is determined by the standard curve external standard method. The content of the target in the cosmetic sample is calculated according to Formula (1). Where: X – the content of the target in the sample, in micrograms per gram (μg/g); ρ – the mass concentration of the target in the sample solution, which is calculated from the standard curve, in micrograms per milliliter (μg/mL); V – the total volume of the tested sample solution, which is converted by the dilution ratio, in milliliters (mL); m – the mass of the weighed sample, in grams (g). ......Source: Above contents are excerpted from the full-copy PDF -- translated/reviewed by: www.ChineseStandard.net / Wayne Zheng et al. 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