Development of pure certified reference material of squalene

In this work, a novel certified reference material (CRM) of squalene (GBW(E) 091179) was developed according to the principles of the ISO Guide. The whole procession including sample preparation, structural analysis, homogeneity assessment, stability study, characterization and uncertainty assessment was studied. The CRM was purified by semi-preparative liquid chromatography on a reversed-phase C18 column from commercial squalene. The certified purity value of the squalene CRM was 99.4 % ± 0.2 % determined by MB and qNMR methods at a confidence level of 95 %(k = 2). [Display omitted] •A novel squalene purity CRM was developed.•Characterization, homogeneity, stability and uncertainty were fully studied.•Mass balance method and qNMR method were used for the characterization of the CRM.•The purity CRM has been approved as reference material GBW(E) 091179. A new certified reference material (CRM) of squalene (GBW(E) 091179) has been developed using a mass balance (MB) approach in conjunction with quantitative nuclear magnetic resonance (qNMR) technique. In compliance with ISO guidelines, the candidate CRM has been investigated for preparation, structure determination, characterization, homogeneity study, stability studies, as well as uncertainty evaluation. The squalene CRM has a certified value of 99.4 %, whose expanded uncertainty is 0.2 % (k = 2). Under a −20 °C condition, the squalene CRM is stable for no less than 12 months, while at 50 °C, its stability duration is 14 days. The uncertainty was evaluated by combining the contributions from characterization, homogeneity, and stability. The developed squalene CRM may be an effective support for method validation, intercomparison in the laboratory, quality control, and metrological traceability used in the food, pharmaceutical, and cosmetic industries.