Vitamin A is a vital nutritional substances that regulates biological activities including development, but is also associated with disease onset. The extent of vitamin A intake influences the retinoid content in the liver, the most important organ for the storage of vitamin A. Measurement of endogenous retinoid in biological samples is important to understand retinoid homeostasis. Here we present a reliable, highly sensitive, and robust method for the quantification of retinol and retinyl palmitate using a reverse-phase HPLC/UV isocratic method. We determined the impact of chronic dietary vitamin A on retinoid levels in livers of mice fed an AIN-93G semi-purified diet (4 IU/g) compared with an excess vitamin A diet (1000 IU/g) over a period from birth to 10 weeks of age. Coefficients of variation for intra-assays for both retinoids were less than 5%, suggesting a higher reproducibility than any other HPLC/UV gradient method. Limits of detection and quantification for retinol were 0.08 pmol, and 0.27 pmol, respectively, which are remarkably higher than previous results. Supplementation with higher doses of vitamin A over the study period significantly increased liver retinol and retinyl palmitate concentrations in adult mice. The assays described here provide a sensitive and rigorous quantification of endogenous retinol and retinyl palmitate, which can be used to help determine retinoid homeostasis in disease states, such as toxic hepatitis and liver cancer.

中文翻译：

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使用液相色谱等度法对视黄醇和棕榈酸视黄酯进行简单而可靠的定量分析

维生素 A 是一种重要的营养物质，可调节包括发育在内的生物活动，但也与疾病发作有关。维生素 A 摄入的程度会影响肝脏中的类维生素 A 含量，肝脏是储存维生素 A 的最重要器官。测量生物样品中的内源性类维生素 A 对于了解类维生素 A 稳态很重要。在这里，我们提出了一种使用反相 HPLC/UV 等度法定量视黄醇和棕榈酸视黄酯的可靠、高灵敏度和稳健的方法。我们确定了长期膳食维生素 A 对喂食 AIN-93G 半纯化饮食（4 IU/g）和过量维生素 A 饮食（1000 IU/g）的小鼠肝脏中维甲酸水平的影响。 10 周龄。两种类视黄醇的测定内变异系数均小于 5%，表明比任何其他 HPLC/UV 梯度方法具有更高的重现性。视黄醇的检测限和定量限分别为 0.08 pmol 和 0.27 pmol，明显高于之前的结果。在研究期间补充更高剂量的维生素 A 可显着增加成年小鼠的肝脏视黄醇和棕榈酸视黄酯浓度。此处描述的测定提供了对内源性视黄醇和视黄醇棕榈酸酯的灵敏和严格的量化，可用于帮助确定疾病状态下的视黄醇稳态，例如中毒性肝炎和肝癌。27 pmol，分别显着高于以前的结果。在研究期间补充更高剂量的维生素 A 可显着增加成年小鼠的肝脏视黄醇和棕榈酸视黄酯浓度。此处描述的测定提供了对内源性视黄醇和视黄醇棕榈酸酯的灵敏和严格的量化，可用于帮助确定疾病状态下的视黄醇稳态，例如中毒性肝炎和肝癌。27 pmol，分别显着高于以前的结果。在研究期间补充更高剂量的维生素 A 可显着增加成年小鼠的肝脏视黄醇和棕榈酸视黄酯浓度。此处描述的测定提供了对内源性视黄醇和视黄醇棕榈酸酯的灵敏和严格的量化，可用于帮助确定疾病状态下的视黄醇稳态，例如中毒性肝炎和肝癌。