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SC/T 3053-2019: Determination of astaxanthin in fish and fishery products by high performance liquid chromatography method Delivery: 9 seconds. True-PDF full-copy in English & invoice will be downloaded + auto-delivered via email. See step-by-step procedure Status: Valid
SC/T 3053-2019: Determination of astaxanthin in fish and fishery products by high performance liquid chromatography method---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/SCT3053-2019 SC AQUACULTURE INDUSTRY STANDARD OF THE PEOPLE’S REPUBLIC OF CHINA ICS 67.120.30 X 20 Determination of Astaxanthin in Fish and Fishery Products by High Performance Liquid Chromatography Method ISSUED ON: AUGUST 01, 2019 IMPLEMENTED ON: NOVEMBER 01, 2019 Issued by: Ministry of Agriculture and Rural Affairs of the People’s Republic of China Table of ContentsForeword ... 3 1 Scope ... 4 2 Normative References ... 4 3 Principle ... 4 4 Reagents ... 4 5 Instruments ... 6 6 Determination Procedures ... 6 7 Calculation of Results ... 8 8 Method Quantification Limit, Recovery Rate and Precision ... 9 Appendix A (Informative) Chromatogram ... 10 Determination of Astaxanthin in Fish and Fishery Products by High Performance Liquid Chromatography Method1 ScopeThis Standard specifies the principle, the used reagents and instruments, the determination procedures, the calculation method of the results, the sensitivity, accuracy and precision of the high-performance liquid chromatography method for the determination of astaxanthin content in fish and fishery products. This Standard is applicable to the determination of astaxanthin content in fish, crustaceans, shrimp meal, krill oil and other products.2 Normative ReferencesThe following documents are essential to the application of this document. For the dated documents, only the versions with the dates indicated are applicable to this document; for the undated documents, only the latest version (including all the amendments) is applicable to this document. GB/T 6682 Water for Analytical Laboratory Use - Specification and Test Methods GB/T 30891 Practice of Sampling Plans for Aquatic Products3 PrincipleThe to-be-tested substance in the sample is extracted by the mixed solution of acetone or dichloromethane-methanol; after alkali saponification, the astaxanthin ester is converted into free astaxanthin; separated by liquid chromatography; measured by ultraviolet detector; and quantified by external standard method.4 Reagents4.1 Unless otherwise specified, the used reagents are all analytically pure; and the water is the Class-I water specified in GB/T 6682. protected from light. The validity period is 1 month. 4.21 Preparation of geometric isomers of astaxanthin: Accurately pipette an appropriate amount of all-trans astaxanthin standard stock solution (4.20); dilute with acetone to prepare a standard solution of 10µg/mL. Pipette 2mL of standard solution into 10mL test tube with a stopper. Add 3mL of dichloromethane; mix well; add 50µL of 0.01 g/mL iodine-dichloromethane solution (4.17); vortex thoroughly; seal and place under natural light for reacting 15min. Then add 1mL of 0.1mol/L sodium thiosulfate solution (4.18) to shake sufficiently and remove excessive iodine; then stand and separate into layers to take the lower phases. After drying with nitrogen, add 1mL of acetone to dissolve it; and it shall be prepared for immediate use. 4.22 Primary secondary amine (PSA) filler: Particle size of 40µm~60µm.5 Instruments5.1 High performance liquid chromatograph: Equipped with ultraviolet detector. 5.2 Analytical balance: Sensitivity of 0.01g. 5.3 Analytical balance: Sensitivity of 0.0001g. 5.4 Analytical balance: Sensitivity of 0.00001g. 5.5 Ultrasonic cleaner. 5.6 Centrifuge: Rotating speed of 8000 r/min. 5.7 Vortex mixer. 5.8 Nitrogen blowing instrument6 Determination Procedures6.1 Preparation of specimen Take a representative specimen and implement it in accordance with the provisions of GB/T 30891. 6.2 Extraction 6.2.1 Fish and crustaceans Take 2g (accurate to 0.01g) of the specimen into a 50mL centrifuge tube; add 4g of anhydrous MgSO4; then add 10mL of acetone; vortex fully. Perform the ultrasonic Where: X - astaxanthin content in the sample, in mg/kg; 1.3 – correction factor of 13-cis astaxanthin against all-trans astaxanthin; A13-cis – peak area of 13-cis astaxanthin in the specimen solution; Atrans – peak area of all-trans astaxanthin in the specimen solution; 1.1 – correction factor of 9-cis astaxanthin against all-trans astaxanthin; A9-cis - peak area of 9-cis astaxanthin in the specimen solution; Cs - all-trans astaxanthin content in the standard working solution, in µg/mL; V – volume of specimen solution, in mL; As – peak area of all-trans astaxanthin standard working solution; m – mass of sample, in g; f – dilution rate.8 Method Quantification Limit, Recovery Rate andPrecision 8.1 Quantification limit The quantification limit of astaxanthin in fish and crustaceans is 2.5mg/kg; the quantification limit of astaxanthin in shrimp meal is 5mg/kg; and the quantification limit of astaxanthin in krill oil is 10mg/kg. 8.2 Recovery rate When the addition concentration of this method is 2.5mg/kg~100mg/kg, the recovery rate shall be 90%~110%. 8.3 Precision The inter-assay coefficient of variation of this method is ≤10%; and the intra-assay coefficient of variation is ≤10%. ......Source: Above contents are excerpted from the full-copy PDF -- translated/reviewed by: www.ChineseStandard.net / Wayne Zheng et al. 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