New Use Agriculture and Natural Plant Products Program, Department of Plant Biology, School of Environmental and Biological Sciences, Rutgers University, 59 Dudley Road, New Brunswick, NJ 08901, USA; Department of Food Science, School of Environmental and Biological Sciences, Rutgers University, 65 Dudley Road, New Brunswick, NJ 08901, USA.
New Use Agriculture and Natural Plant Products Program, Department of Plant Biology, School of Environmental and Biological Sciences, Rutgers University, 59 Dudley Road, New Brunswick, NJ 08901, USA.
J Chromatogr B Analyt Technol Biomed Life Sci. 2020 Jul 15;1149:122146. doi: 10.1016/j.jchromb.2020.122146. Epub 2020 May 19.
Raspberry ketone (RK) (4-(4-hydroxyphenyl)-2-butanone) is the major compound responsible for the characteristic aroma of red raspberries, and has long been used commercially as a flavoring agent and recently as a weight loss supplement. A targeted UHPLC-QqQ-MS/MS method was developed and validated for analysis of RK and 25 associated metabolites in mouse plasma and brain. Dispersion and projection analysis and central composite design were used for method optimization. Random effect analysis of variance was applied for validation inference and variation partition. Within this framework, repeatability, a broader sense of precision, was calculated as fraction of accuracy variance, reflecting instrumental imprecision, compound degradation and carry-over effects. Multivariate correlation analysis and principle component analysis were conducted, revealing underlying association among the manifold of method traits. R programming was engaged in streamlined statistical analysis and data visualization. Two particular phenomena, the analytes' background existence in the enzyme solution used for phase II metabolites deconjugation, and the noted lability of analytes in pure solvent at 4 ℃ vs. elevated stability in biomatrices, were found critical to method development and validation. The approach for the method development and validation provided a foundation for experiments that examine RK metabolism and bioavailability.
覆盆子酮（RK）（4-（4-羟苯基）-2-丁酮）是赋予红覆盆子特有香气的主要化合物，长期以来一直被用作商业香料，并最近被用作减肥补充剂。开发并验证了一种靶向 UHPLC-QqQ-MS/MS 方法，用于分析小鼠血浆和脑中的 RK 和 25 种相关代谢物。分散和投影分析以及中心复合设计用于方法优化。随机效应方差分析用于验证推断和变异划分。在此框架内，重复性，更广泛的精密度概念，计算为准确度方差的分数，反映仪器不精密度，化合物降解和交叉污染效应。进行了多元相关分析和主成分分析，揭示了方法特征之间的潜在关联。R 编程用于简化统计分析和数据可视化。发现了两个特殊现象，用于 II 期代谢物去共轭的酶溶液中分析物的背景存在，以及在 4℃下纯溶剂中分析物的明显不稳定性与生物基质中更高的稳定性之间的关联，这对方法开发和验证至关重要。该方法开发和验证方法为研究 RK 代谢和生物利用度的实验提供了基础。
