Methods
Methods
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DOI：
10.1111/j.1651-2227.1944.tb10559.x
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发表时间：
1944-12
期刊：
影响因子：
--
通讯作者：
James A. Gott
中科院分区：
文献类型：
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作者：
James A. Gott
目的：覆盆子酮（4-（4-羟基苯基）-2-丁酮）（RK）是红覆盆子的主要香气化合物，在商业上用作调味剂。它现在作为减肥补充剂而受到关注。本工作旨在建立一种靶向超高效液相色谱三重四极杆串联质谱（UHPLC-QqQ-MS/MS）方法来分析小鼠血浆和脑中的覆盆子酮及其25种相关代谢物。方法：采用色散投影分析和中心组合设计对LC-MS方法进行优化。应用方差随机效应分析来验证推断和变异划分。在此框架内，可重复性（更广泛的精度概念）被计算为精度方差的分数，反映了仪器的不精确性、化合物降解和残留效应。进行了多变量相关分析和主成分分析，揭示了多种方法特征之间潜在的相互联系和关联。 R 编程致力于简化统计分析和数据可视化。结果：所开发并验证的 RK 和 25 种代谢物方法的准确性、基质效应、回收率和处理效率大多在 80∼120% 范围内，精密误差低于 5%，定量限为 0.4∼6.0 ng/mL，线性范围跨越三个数量级。我们发现两个特殊现象对于方法开发和验证至关重要，即用于 II 相代谢物解偶联的酶溶液中分析物的背景存在，以及 4 °C 纯溶剂中分析物相对于生物基质中稳定性提高的明显责任。结论：开发并验证的 UHPLC-QqQ-MS/MS 方法为进一步研究 RK 代谢和生物利用度提供了重要工具。资金来源：国立卫生研究院。
Objectives: Raspberry ketone (4-(4-hydroxyphenyl)-2-butanone) (RK) is the major aroma compound of red raspberries and used commercially as flavoring agent. It is now gaining attention as a weight loss supplement. This work aimed to establish atargeted ultrahigh performance liquid chromatography triple quadrupole tandem mass spectrometry (UHPLC-QqQ-MS/MS) method for analysis of raspberry ketone and its 25 associatedmetabolites inmouse plasma and brain. Methods:Dispersion and projection analysis and central composite design were used for LC-MS 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 interlinkage and association among the manifold of method traits. R programming was engaged in streamlined statistical analysis and data visualization. Results: The developed and validated method for RK and 25 metabolites had accuracy, matrix effect, recovery and processing efficiency bound mostly within the 80∼120% range, precision error below 5%, limit of quantification at 0.4∼6.0 ng/mL and linear range across three orders of magnitude. Two particular phenomena, the analytes’ background existence in the enzyme solution used for phase II metabolites deconjugation, and the noted liability of analytes in pure solvent at 4 °C vs. elevated stability in biomatrices, were found critical to method development and validation. Conclusions: The UHPLC-QqQ-MS/MS method developed and validated provided an important tool for further investigation of RK metabolism and bioavailability. Funding Sources: National Institutes of Health.