MICROSCALE SERUM EXTRACTION METHOD FOR THE SIMULTANEOUS ANALYSIS OF CORTICOSTERONE AND LIPIDS.
MICROSCALE SERUM EXTRACTION METHOD FOR THE SIMULTANEOUS ANALYSIS OF CORTICOSTERONE AND LIPIDS.
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DOI：
10.1039/c9ay01757g
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发表时间：
2019-11
期刊：
影响因子：
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通讯作者：
Alana L Rister;K. Bidne;J. Wood;Eric D. Dodds
中科院分区：
文献类型：
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作者：
Alana L Rister;K. Bidne;J. Wood;Eric D. Dodds
皮质酮是调节代谢和应激反应的重要类固醇。现有的皮质酮测定方法包括放射免疫法（RIA）、酶联免疫吸附法（ELISA）和液相色谱-质谱法（LC-MS）。虽然每种方法都有其优点，但ria使用的放射性同位素需要特别规范的使用和处置。此外，elisa和ria都需要昂贵的试剂盒，并且只能测量单一分析物。在这项研究中，我们建立了一种基于改良的Folch提取的新样品制备技术，该技术允许同时从血清中分离皮质酮和脂质。然后用LC-MS对提取物进行分析。仅使用5µL血清，皮质酮的定量变异系数为3%或更低，检测限为0.12µM。总的来说，这项研究的结果应该有利于使用小体积样本的各种研究中循环皮质酮和脂质的测量。
Corticosterone is an important steroid for the regulation of metabolism and stress response. Existing methods for the measurement of corticosterone include radioimmunoassay (RIA), enzyme linked immunosorbent assay (ELISA), and liquid chromatography-mass spectrometry (LC-MS). While each of these approaches have their advantages, RIAs use radioactive isotopes that necessitate specially regulated usage and disposal. Furthermore, both ELISAs and RIAs require expensive kits and can only measure a single analyte. In this study, we establish a new sample preparation technique based on a modified Folch extraction that allows for the simultaneous isolation of corticosterone and lipids from serum. The extract is then analyzed by LC-MS. Using only 5 µL of serum, quantification of corticosterone was achieved with coefficients of variation at 3% or less and a detection limit of 0.12 µM. Overall, the results of this study should be beneficial to the measurement of circulating corticosterone and lipids for a variety of studies using small volumes of samples.