Quantification of endogenous retinoic acid in limited biological samples by LC/MS/MS
Quantification of endogenous retinoic acid in limited biological samples by LC/MS/MS
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DOI：
10.1042/bj20041867
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发表时间：
2005-05-15
影响因子：
4.1
通讯作者：
Napoli, JL
中科院分区：
文献类型：
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作者：
Kane, MA;Chen, N;Napoli, JL
我们报告了一种灵敏的LC（液相色谱）/MS/MS测定法，使用选择性反应监测来定量RA（维甲酸），该方法适用于有限大小（10 - 20 mg组织湿重）的生物样品，不需要样品衍生化，提供质量鉴定并将atRA（全反式RA）从其几何异构体中分离出来。该测定在20 fmol至10 pmol的线性范围内定量，在信噪比为3时的检测限为10 fmol。变异系数为：仪器，0.5 - 2.9%;批内，5.4 +/-0.4%;批间8.9 +/-1.0%。内标（全反式-4，4-二甲基-RA）通过确认提取效率和揭示处理诱导的异构化来提高准确度。2 - 4月龄C57 BL/6雄性小鼠的组织中atRA浓度为7 - 9.6 pmol/g，血清atRA浓度为1.9 +/-0.6 pmol/ml（+/-S.E.M.）。组织13-cis-RA范围为2.9 - 4.2 pmol/g，血清13-cis-RA为1.2 +/-0.3 pmol/ml。CRBP（细胞视黄醇结合蛋白）缺失小鼠肝脏的atRA比野生型低30%（P
We report a sensitive LC (liquid chromatography)/MS/MS assay using selected reaction monitoring to quantify RA (retinoic acid), which is applicable to biological samples of limited size (10-20 mg of tissue wet weight), requires no sample derivatization, provides mass identification and resolves atRA (all-trans-RA) from its geometric isomers. The assay quantifies over a linear range of 20 fmol to 10 pmol, and has a 10 fmol limit of detection at a signal/noise ratio of 3. Coefficients of variation are: instrumental, 0.5-2.9%; intra-assay, 5.4 +/- 0.4%; inter-assay 8.9 +/- 1.0%. An internal standard (all-trans-4,4-dimethyl-RA) improves accuracy by confirming extraction efficiency and revealing handling-induced isomerization. Tissues of 2-4-month-old C57BL/6 male mice had atRA concentrations of 7-9.6 pmol/g and serum atRA of 1.9 +/- 0.6 pmol/ml (+/- S.E.M.). Tissue 13-cis-RA ranged from 2.9 to 4.2 pmol/g, and serum 13-cis-RA was 1.2 +/- 0.3 pmol/ml. CRBP (cellular retinol-binding protein)-null mouse liver had atRA similar to 30% lower than wild-type (P