Quantitation of retinaldehyde in small biological samples using ultrahigh-performance liquid chromatography tandem mass spectrometry.
Quantitation of retinaldehyde in small biological samples using ultrahigh-performance liquid chromatography tandem mass spectrometry.
复制标题
DOI：
10.1016/j.ab.2015.05.016
复制
发表时间：
2015-09-01
影响因子：
2.9
通讯作者：
Napoli JL
中科院分区：
文献类型：
--
作者：
Wang J;Yoo HS;Obrochta KM;Huang P;Napoli JL
我们报告了一种UHPLC-MS/MS来定量有限大小（15-35 mg）的生物样品中的全反式视黄醇，这对于与脂肪一起使用特别有利。为了便于回收，将视黄醛和内标物3，4-二脱氢视黄醛原位衍生成它们的O-乙基肟。UHPLC分辨率与MS/MS的高灵敏度和特异性相结合，可以定量测定视黄醇-O-乙基肟，检测下限为5 fmol，线性范围为5 fmol至1 pmol。该测定显示，多种组织中的视网膜的眼外浓度范围为约2至40 pmol/g：与全反式维甲酸的范围相同。全反式维甲酸对其核受体（RARα、β、γ）具有高亲和力（kd ≤ 0.4 nM），而视黄醛对其核受体（RARα、β、γ）的亲和力较低（如果有），因此这些浓度的视黄醛不太可能激活RAR。我们还表明，大量的维生素A在食物中使用的饮食增加视网膜的脂肪贮库2至5倍，相对于水平的脂肪喂养的维生素A充足的饮食，建议为实验室啮齿动物。该测定法也可用于定量体外视黄醇转化为视黄醛的转化，因此提供了一种有效的体内和体外研究视黄醇代谢的方法。
We report an UHPLC-MS/MS to quantify all-trans-retinal in biological samples of limited size (15-35 mg), which is especially advantageous for use with adipose. To facilitate recovery, retinal and the internal standard 3,4-didehydroretinal were derivatized in situ into their O-ethyloximes. UHPLC resolution combined with high sensitivity and specificity of MS/MS allowed quantification of retinal-O-ethyloximes with a 5 fmol lower limit of detection and a linear range from 5 fmol to 1 pmol. This assay revealed that extra-ocular concentrations of retinal range from ~2 to 40 pmol/g in multiple tissues: the same range as all-trans-retinoic acid. All-trans-retinoic acid has high affinity (kd ≤ 0.4 nM) for its nuclear receptors (RARα, β, γ), whereas retinal has low, if any affinity for these receptors, making it unlikely that these retinal concentrations would activate RAR. We also show that the copious amount of vitamin A used in chow diets increases retinal in adipose depots 2 to 5-fold relative to levels in adipose of mice fed a vitamin A-sufficient diet, as recommended for laboratory rodents. This assay also is proficient for quantifying conversion of retinol into retinal in vitro, and therefore provides an efficient method to study metabolism of retinol in vivo and in vitro.
登录
查看更多内容
影响因子：
3.2
作者：
Foster, RG;Provencio, I;DeGrip, WJ
通讯作者：
DeGrip, WJ
DOI：
10.1016/s0021-9673(00)90269-3
发表时间：
1988-04-22
期刊：
JOURNAL OF CHROMATOGRAPHY
影响因子：
--
作者：
LANDERS, GM;OLSON, JA
通讯作者：
OLSON, JA
影响因子：
4.8
作者：
Eroglu, Abdulkerim;Hruszkewycz, Damian P.;Harrison, Earl H.
通讯作者：
Harrison, Earl H.
影响因子：
4.1
作者：
Kane, MA;Chen, N;Napoli, JL
通讯作者：
Napoli, JL
影响因子：
2.9
作者：
Schmidt, CK;Brouwer, A;Nau, H
通讯作者：
Nau, H