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首页> 外文期刊>Analytical and bioanalytical chemistry >Application of dried spot cards as a rapid sample treatment method for determining hydroxytyrosol metabolites in human urine samples. Comparison with microelution solid-phase extraction
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Application of dried spot cards as a rapid sample treatment method for determining hydroxytyrosol metabolites in human urine samples. Comparison with microelution solid-phase extraction

机译：干点卡作为一种快速样品处理方法，用于测定人尿液样品中的羟基酪醇代谢物。与微洗脱固相萃取的比较

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摘要

Two different rapid sample pretreatment strategies, dried spot cards, and microelution solid-phase extraction plates (μSPE), with ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS) have been developed and validated for the determination of hydroxytyrosol and its metabolites in spiked human urine samples. Hydroxytyrosol, hydroxytyrosol-3'-O-glucuronide, hydroxytyrosol-4'- O-glucuronide, hydroxytyrosol-3-O-sulphate, and homovanillic alcohol-4'-O-glucuronide were used as the target compounds. Using the FTA DMPK-A dried urine spot card under optimum conditions, with 5 μL of preconcentrated urine volume and 100 μL of methanol/water (50/50, v/v) as the elution solvent, the extraction recovery (%R) of the compounds studied was higher than 80 %, and the matrix effect (%ME) was less than 8 %. The stability of these cards and punching at the centre or side of the card were also studied, obtaining an excellent stability after 7 days of storage and complete homogeneity across the surface of the dried drop. The different μSPE parameters that affect the efficiency were also studied, and under optimum conditions, the %R and the %ME were higher than 70 % and lower than 17 %, respectively. The linearity range in dried urine spot cards was 2.5-20 μM for all the metabolites, with the exception of hydroxytyrosol-3-O-sulphate and hydroxytyrosol, which were 0.3-70 μM and 2.5-50 μM respectively. With regards to μSPE, the linearity range was 0.5-5 μM for all the studied compounds, except for hydroxytyrosol-3-O-sulphate, which was 0.08-5 μM. The quantification limits (LOQs) were 0.3-2.5 μM and 0.08-0.5 μM in dried spot cards and in μSPE, respectively. The two developed methods were then applied and compared for determining hydroxytyrosol and its metabolites in human 24 h-urine samples after a sustained consumption (21 days) of a phenol-enriched virgin olive oil. The metabolites identified were hydroxytyrosol in its glucuronide and sulphate forms, homovanillic alcohol in its glucuronide and sulphate forms, homovanillic acid sulphate and hydroxytyrosol acetate sulphate.
机译：已经开发了两种不同的快速样品预处理策略，干点卡和微洗脱固相萃取板（μSPE），以及超高效液相色谱-串联质谱（UPLC-MS / MS），可用于测定加标的人类尿液样品中的羟基酪醇及其代谢物。使用羟基酪醇，羟基酪醇-3'-O-葡糖醛酸，羟基酪醇-4'-O-葡糖醛酸，羟基酪醇-3-O-硫酸盐和高香草醇-4'-O-葡糖醛酸作为目标化合物。使用FTA DMPK-A干尿斑点卡在最佳条件下，以5μL的预浓缩尿液体积和100μL的甲醇/水（50/50，v / v）作为洗脱溶剂，提取的回收率（％R）研究的化合物含量高于80％，基质效应（％ME）低于8％。还研究了这些卡片的稳定性以及在卡片中央或侧面的打孔，在储存7天后具有出色的稳定性，并且在干燥液滴的整个表面上完全均匀。还研究了影响效率的不同μSPE参数，在最佳条件下，％R和％ME分别高于70％和低于17％。干燥尿斑卡中所有代谢物的线性范围均为2.5-20μM，除了羟基酪醇-3-O-硫酸盐和羟基酪醇分别为0.3-70μM和2.5-50μM。关于μSPE，所有研究化合物的线性范围为0.5-5μM，除了羟基酪醇-3-O-硫酸盐为0.08-5μM。干燥斑点卡和μSPE中的定量限（LOQ）分别为0.3-2.5μM和0.08-0.5μM。然后应用这两种已开发的方法，并比较了在持续食用（富含21天）富含苯酚的初榨橄榄油后，人24小时尿液样本中的羟基酪醇及其代谢物的含量。鉴定出的代谢产物是葡萄糖醛酸和硫酸盐形式的羟基酪醇，葡萄糖醛酸和硫酸盐形式的高香草醇，硫酸高香草酸和乙酸羟基酪醇硫酸盐。

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