Vitamin E deficiencies can impact normal growth and development in humans and animals, and assessment of circulating levels of vitamin E and its metabolites may be an important endpoint for evaluation. Development of a sensitive method to detect and quantify low concentrations of vitamin E and metabolites in biological specimens allows for a proper diagnosis for patients and animals that are deficient. We developed a method to simultaneously extract, detect, and quantify the vitamin E compounds alpha-tocopherol (α-TP), gamma-tocopherol (γ-TP), alpha-tocotrienol (α-TT), and gamma-tocotrienol (γ-TT), and the corresponding metabolites formed after β-oxidation of α-TP and γ-TP, alpha-carboxymethylbutyl hydroxychroman (α-CMBHC) and alpha- or gamma-carboxyethyl hydroxychroman (α- or γ-CEHC), respectively, from equine plasma and serum. Quantification was achieved through liquid chromatography–tandem mass spectrometry. We applied a 96-well high-throughput format using a Phenomenex Phree plate to analyze plasma and serum. Compounds were separated by using a Waters ACQUITY UPLC BEH C18 column with a reverse-phase gradient. The limits of detection for the metabolites and vitamin E compounds were 8–330 pg/mL. To validate the method, intra-day and inter-day accuracy and precision were evaluated along with limits of detection and quantification. The method was then applied to determine concentrations of these analytes in plasma and serum of horses. Alpha-TP levels were 3–6 µg/mL of matrix; the metabolites were found at much lower levels, 0.2–1.0 ng/mL of matrix.

维生素 E 缺乏会影响人类和动物的正常生长和发育，评估维生素 E 及其代谢物的循环水平可能是评估的重要终点。开发一种灵敏的方法来检测和量化生物样本中低浓度的维生素 E 和代谢物，可以对缺乏维生素 E 的患者和动物进行适当的诊断。我们开发了一种同时提取、检测和量化维生素 E 化合物 α-生育酚 (α-TP)、γ-生育酚 (γ-TP)、α-生育三烯酚 (α-TT) 和 γ-生育三烯酚 (γ- TT)，以及分别来自马血浆和血清。通过液相色谱-串联质谱法进行定量。我们使用 Phenomenex Phree 板应用 96 孔高通量形式来分析血浆和血清。使用具有反相梯度的 Waters ACQUITY UPLC BEH C18 色谱柱分离化合物。代谢物和维生素 E 化合物的检测限为 8–330 pg/mL。为了验证该方法，对日内和日间准确度和精密度以及检测限和定量限进行了评估。然后应用该方法确定这些分析物在马血浆和血清中的浓度。Alpha-TP 水平为 3–6 µg/mL 基质；代谢物的含量要低得多，为 0.2–1.0 ng/mL 基质。我们使用 Phenomenex Phree 板应用 96 孔高通量形式来分析血浆和血清。使用具有反相梯度的 Waters ACQUITY UPLC BEH C18 色谱柱分离化合物。代谢物和维生素 E 化合物的检测限为 8–330 pg/mL。为了验证该方法，对日内和日间准确度和精密度以及检测限和定量限进行了评估。然后应用该方法确定这些分析物在马血浆和血清中的浓度。Alpha-TP 水平为 3–6 µg/mL 基质；代谢物的含量要低得多，为 0.2–1.0 ng/mL 基质。我们使用 Phenomenex Phree 板应用 96 孔高通量形式来分析血浆和血清。使用具有反相梯度的 Waters ACQUITY UPLC BEH C18 色谱柱分离化合物。代谢物和维生素 E 化合物的检测限为 8–330 pg/mL。为了验证该方法，对日内和日间准确度和精密度以及检测限和定量限进行了评估。然后应用该方法确定这些分析物在马血浆和血清中的浓度。Alpha-TP 水平为 3–6 µg/mL 基质；代谢物的含量要低得多，为 0.2–1.0 ng/mL 基质。代谢物和维生素 E 化合物的检测限为 8–330 pg/mL。为了验证该方法，对日内和日间准确度和精密度以及检测限和定量限进行了评估。然后应用该方法确定这些分析物在马血浆和血清中的浓度。Alpha-TP 水平为 3–6 µg/mL 基质；代谢物的含量要低得多，为 0.2–1.0 ng/mL 基质。代谢物和维生素 E 化合物的检测限为 8–330 pg/mL。为了验证该方法，对日内和日间准确度和精密度以及检测限和定量限进行了评估。然后应用该方法确定这些分析物在马血浆和血清中的浓度。Alpha-TP 水平为 3–6 µg/mL 基质；代谢物的含量要低得多，为 0.2–1.0 ng/mL 基质。