Shirota Functional Foods, S.L., Carrer de Mèxic, Reus, Spain.
J Sep Sci. 2010 Sep;33(17-18):2841-53. doi: 10.1002/jssc.201000274.
In the analysis of biological samples, such as plasma or serum, the quantity of sample available is a critical parameter in most cases. A good approach is the use of the microelution SPE (μSPE) plates as sample pre-treatment technique in which the loaded sample volume is low. An off-line μSPE and ultra-performance LC-ESI-MS/MS (UPLC-ESI-MS/MS) method was developed and validated to determine procyanidins and anthocyanins in spiked plasma samples. The sample pre-treatment μSPE allowed the simultaneous determination of procyanidins and anthocyanins from plasma by using a small sample volume (350 μL) and without an evaporation step previous to the chromatographic analysis. Moreover, the use of UPLC technique allowed to determine the studied compounds at low concentration levels in a short analysis time (12.5 min approximately). Then, the developed method was applied to determine the studied compounds, procyanidins and anthocyanins, and their metabolites in rat plasma samples. Previously, the rats had consumed 5000 mg/kg of a grape pomace extract and the plasma was extracted 4 h after administration. The procyanidins catechin and epicatechin glucuronide, methyl catechin and epicatechin glucuronide, and methyl catechin and epicatechin sulphate were detected at μM concentration level, and the parent anthocyanins at nM.
在分析生物样本（如血浆或血清）时，大多数情况下，可用样本的数量是一个关键参数。一种好的方法是使用微洗脱 SPE（μSPE）板作为样品预处理技术，其中加载的样品体积较低。开发并验证了一种离线 μSPE 和超高效 LC-ESI-MS/MS（UPLC-ESI-MS/MS）方法，用于测定血浆中前花青素和花色苷的含量。该样品预处理 μSPE 允许使用小体积（350 μL）且在色谱分析之前无需蒸发步骤同时从血浆中测定前花青素和花色苷。此外，使用 UPLC 技术可在短分析时间（约 12.5 分钟）内测定低浓度水平下研究的化合物。然后，该方法应用于测定大鼠血浆样品中研究的化合物、前花青素和花色苷及其代谢物。在此之前，大鼠已经消耗了 5000 mg/kg 的葡萄渣提取物，并且在给药后 4 小时提取血浆。以 μM 浓度水平检测到原花青素儿茶素和表儿茶素葡萄糖苷、甲基儿茶素和表儿茶素葡萄糖苷、以及甲基儿茶素和表儿茶素硫酸盐，而母体花色苷则以 nM 浓度水平检测到。
