Fang Cheng, Zhuang Xiaotong, Li Zhanguo, Zou Yongfang, Pu Jizhou, Wang Dong, Xu Yan
Laboratory of Brewing Microbiology and Applied Enzymology, School of Biotechnology, Jiangnan University, Wuxi 214122, China.
Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China.
Metabolites. 2025 Apr 2;15(4):246. doi: 10.3390/metabo15040246.
: Post-production storage plays a pivotal role in developing the characteristic flavor profile of , a traditional alcoholic beverage in China. While aging markers remain crucial for quality authentication, the identification of reliable metabolic indicators for chronological determination requires further exploration. : This study establishes a novel liquid chromatography-tandem mass spectrometry (LC-MS/MS) methodology for quantifying five linoleic acid-derived oxidative metabolites in : 9,12,13-trihydroxy-10(E)-octadecenoic acid (9,12,13-TriHOME), 9,10-Dihydroxy-12-octadecenoic acid (9,10-DiHOME), 9-oxo-(10E,12Z)-octadecadienoic acid (9-OxoODE), 9-hydroxy-(10E,12Z)-octadecadienoic acid (9-HODE) and 13-hydroxyoctadeca-(9Z,11E)-octadecadienoic acid (13-HODE). : The optimized protocol demonstrated exceptional sensitivity with limits of detection at 0.4 ppb through membrane-filtered direct dilution. Calibration curves exhibited excellent linearity (R > 0.9990) across 1.0-100.0 ppb ranges. Method validation revealed satisfactory recovery rates (87.25-119.44%) at three spiking levels (10/20/50 ppb) with precision below 6.96% RSD. Application to authentic samples showed distinct temporal accumulation patterns. Light-aroma exhibited storage duration-dependent increases in all five oxides. Strong aroma variants demonstrated significant positive correlations for 9,12,13-TriHOME, 9,10-DiHOME, and 9-OxoODE with aging time. : These findings systematically characterize linoleic acid oxidation products as potential aging markers, providing both methodological advancements and new insights into aging mechanisms.
后熟储存对于中国传统酒精饮料[具体酒名未给出]特征风味的形成起着关键作用。虽然陈酿标记对于质量认证仍然至关重要,但确定可靠的代谢指标用于年代测定还需要进一步探索。本研究建立了一种新颖的液相色谱 - 串联质谱(LC - MS/MS)方法,用于定量[具体酒名未给出]中五种亚油酸衍生的氧化代谢物:9,12,13 - 三羟基 - 10(E) - 十八碳烯酸(9,12,13 - TriHOME)、9,10 - 二羟基 - 12 - 十八碳烯酸(9,10 - DiHOME)、9 - 氧代 - (10E,12Z) - 十八碳二烯酸(9 - OxoODE)、9 - 羟基 - (10E,12Z) - 十八碳二烯酸(9 - HODE)和13 - 羟基十八碳 - (9Z,11E) - 十八碳二烯酸(13 - HODE)。优化后的方案具有出色的灵敏度,通过膜过滤直接稀释,检测限为0.4 ppb。校准曲线在1.0 - 100.0 ppb范围内呈现出优异的线性(R > 0.9990)。方法验证表明,在三个加标水平(10/20/50 ppb)下回收率令人满意(87.25 - 119.44%),精密度低于6.96% RSD。应用于真实样品显示出明显的时间积累模式。清香型[具体酒名未给出]中所有五种氧化物的含量均随储存时间增加。浓香型变体中,9,12,13 - TriHOME、9,10 - DiHOME和9 - OxoODE与陈酿时间呈显著正相关。这些发现系统地将亚油酸氧化产物表征为潜在的陈酿标记,既提供了方法学上的进步,也为[具体酒名未给出]的陈酿机制提供了新的见解。