HPLC/UV quantitation of retinal, retinol, and retinyl esters in serum and tissues
HPLC/UV quantitation of retinal, retinol, and retinyl esters in serum and tissues
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DOI：
10.1016/j.ab.2008.03.038
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发表时间：
2008-07-01
影响因子：
2.9
通讯作者：
Napoli, Joseph L.
中科院分区：
文献类型：
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作者：
Kane, Maureen A.;Folias, Alexandra E.;Napoli, Joseph L.
我们报告了用于定量视黄酯（RE）、视黄醇（ROL）和视网膜（RAL）的稳健HPLC/UV方法，适用于不同的生物样品，检测下限分别为0.7、0.2和0.2 pmol，线性范围大于3个数量级。这些测定对小的复杂生物样品（10-20 mg组织）具有良好的功能。变异系数范围为5.9 - 10.0%（日内）和5.9 - 11.0%（日间）。小鼠血清和组织（肝、肾、脂肪、肌肉、脾、睾丸、皮肤、脑和脑区域）中内源性RE、ROL和RAL的定量揭示了实用性。用C57 BL/6小鼠脑基因座（海马、皮质、嗅球、丘脑、小脑和纹状体）说明了区分ROL和RE的空间浓度的能力。我们还开发了一种方法来区分异构形式的ROL调查前体的视黄酸。ROL异构体测定的检测限在3.5和4.5 proof之间，并且具有与RCL/RE和RAL测定相似的线性范围和变异系数。本文所述的测定提供了内源性RE、ROL和RAL的灵敏和严格的定量，以阐明疾病状态如阿尔茨海默病、2型糖尿病、肥胖症和癌症中的类维生素A稳态。(c)2008年爱思唯尔公司All rights reserved.
We report robust HPLC/UV methods for quantifying retinyl esters (RE), retinol (ROL), and retinal (RAL) applicable to diverse biological samples with lower limits of detection of 0.7, 0.2, and 0.2 pmol, respectively, and linear ranges greater than 3 orders of magnitude. These assays function well with small, complex biological samples (10-20 mg tissue). Coefficients of variation range from 5.9 to 10.0% (intraday) and from 5.9 to 11.0% (interday). Quantification of endogenous RE, ROL, and RAL in mouse serum and tissues (liver, kidney, adipose, muscle, spleen, testis, skin, brain, and brain regions) reveals utility. Ability to discriminate spatial concentrations of ROL and RE is illustrated with C57BL/6 mouse brain loci (hippocampus, cortex, olfactory bulb, thalamus, cerebellum, and striatum). We also developed a method to distinguish isomeric forms of ROL to investigate precursors of retinoic acid. The ROL isomer assay has limits of detection between 3.5 and 4.5 proof and has a linear range and coefficient of variation similar to those of the RCL/RE and RAL assays. The assays described here provide for sensitive and rigorous quantification of endogenous RE, ROL, and RAL to elucidate retinoid homeostasis in disease states such as Alzheimer's disease, type 2 diabetes, obesity, and cancer. (c) 2008 Elsevier Inc. All rights reserved.