Avula Bharathi, Parveen Iffat, Zhao Jianping, Wang Mei, Techen Natascha, Wang Yan-Hong, Riaz Mohammad, Bae Ji-Yeong, Shami Anter A, Chittiboyina Amar G, Khan Ikhlas A, Sharp Joshua S
National Center for Natural Products Research, School of Pharmacy, University of Mississippi, University, MS, USA.
Department of BioMolecular Sciences, School of Pharmacy, University of Mississippi, MS, USA University.
J Diet Suppl. 2022;19(4):515-533. doi: 10.1080/19390211.2021.1897724. Epub 2021 Mar 25.
The presence of bio-macromolecules as major ingredients is a primary factor in marketing many biologically derived macromolecular supplements. Workflows for analyzing these supplements for quality assurance, adulteration, and other supply-chain difficulties must include a qualitative assessment of small-molecule and macromolecular components; however, no such integrated protocol has been reported for these bio-macromolecular supplements. Twenty whey protein supplements were analyzed using an integrated workflow to identify protein content, protein adulteration, inorganic elemental content, and macromolecular and small-molecule profiles. Orthogonal analytical methods were employed, including NMR profiling, LC-DAD-QToF analysis of small-molecule components, ICP-MS analysis of inorganic elements, determination of total protein content by a Bradford assay, SDS-PAGE protein profiling, and bottom-up shotgun proteomic analysis using LC-MS-MS. All 20 supplements showed a reduced protein content compared to the claimed content but no evidence of adulteration with protein from an unclaimed source. Many supplements included unlabeled small-molecule additives (but nontoxic) and significant deviations in metal content, highlighting the importance of both macromolecular and small-molecule analysis in the comprehensive profiling of macromolecular supplements. An orthogonal, integrated workflow allowed the detection of crucial product characteristics that would have remained unidentified using traditional workflows involving either analysis of small-molecule nutritional supplements or protein analysis.
生物大分子作为主要成分的存在是许多生物衍生大分子补充剂营销的一个主要因素。用于分析这些补充剂的质量保证、掺假和其他供应链难题的工作流程必须包括对小分子和大分子成分的定性评估；然而，尚未有针对这些生物大分子补充剂的此类综合方案的报道。使用综合工作流程对20种乳清蛋白补充剂进行了分析，以确定蛋白质含量、蛋白质掺假、无机元素含量以及大分子和小分子概况。采用了正交分析方法，包括核磁共振波谱分析、小分子成分的液相色谱-二极管阵列-四极杆飞行时间质谱分析、无机元素的电感耦合等离子体质谱分析、考马斯亮蓝法测定总蛋白含量、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳蛋白质分析以及使用液相色谱-串联质谱的自下而上鸟枪法蛋白质组学分析。所有20种补充剂的蛋白质含量均低于宣称含量，但没有证据表明掺有未宣称来源的蛋白质。许多补充剂含有未标明的小分子添加剂（但无毒）且金属含量存在显著偏差，这突出了大分子和小分子分析在大分子补充剂综合分析中的重要性。一种正交的综合工作流程能够检测到关键的产品特性，而使用涉及小分子营养补充剂分析或蛋白质分析的传统工作流程这些特性将无法被识别。
