Rapid determination of corticosterone in mouse plasma by ultra fast liquid chromatography-tandem mass spectrometry
Rapid determination of corticosterone in mouse plasma by ultra fast liquid chromatography-tandem mass spectrometry
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超快速液相色谱-串联质谱法快速测定小鼠血浆中的皮质酮。
DOI：
10.1002/bmc.3232
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发表时间：
2014-12-01
影响因子：
1.8
通讯作者：
Cai, Bao-Chang
中科院分区：
文献类型：
--
作者：
Li, Huan;Liu, Xiao;Cai, Bao-Chang
重度抑郁症是一种严重的、威胁生命的、高度流行的精神疾病。很大一部分抑郁症患者的特征是下丘脑-垂体-肾上腺轴活动过度，导致血浆糖皮质激素（人的皮质醇和啮齿动物的皮质酮）升高。糖皮质激素是抑郁症发病的关键分子。首次建立了一种简单、灵敏度高、特异性强的超快速液相色谱-串联质谱法测定小鼠血浆中皮质酮的方法，为生物样品中皮质酮的临床高通量测定提供了技术支持。样品加标甲醇以沉淀蛋白质，然后在Agilent Zorbax Eclipse Plus C18（100 × 2.1 mm，1.8 µm）色谱柱上进行色谱分析，以甲醇和0.1%甲酸作为移动的相，在5 min内进行线性梯度洗脱。采用超快速液相色谱-三重四极杆串联质谱法在正离子中检测皮质酮。皮质酮和氢化可的松（内标）的离子[M + H]（+）m/z 347.2 → m/z 311.1和[M + H]（+）m/z 363.2 → m/z 327.2用于定量。皮质酮的定量下限为1 ng/mL。该方法已成功应用于小鼠血浆中皮质酮的定量测定。
Major depressive disorder is a severe, life-threatening and highly prevalent psychiatric disorder. A high percentage of people suffering from depression are characterized by hyperactivity of the hypothalamic-pituitary-adrenal axis, resulting in plasma glucocorticoid (cortisol in human and corticosterone in rodent) elevations. Glucocorticoid is a critical molecule in the onset of pathology of depression. A simple, highly sensitive and specific method based on ultra-fast liquid chromatography-tandem mass spectrometry method has been developed for the quantitation of corticosterone in mouse plasma for the first time, which provides technical support for the high-throughput measurement for clinical determination of corticosterone in biological samples. Samples were spiked with methanol to precipitate the protein, and then chromatographed on an Agilent Zorbax Eclipse Plus C18 (100 × 2.1 mm,1.8 µm) column by linear gradient elution with methanol and 0.1% formic acid as the mobile phase within 5 min. The detection of corticosterone was performed on ultra-fast liquid chromatography-triple quadrupole tandem mass spectrometry in the positive ion. The ions [M + H](+) m/z 347.2 → m/z 311.1 for corticosterone and [M + H](+) m/z 363.2 → m/z 327.2 for hydrocortisone (internal standard) were used for quantitative determination. The lower quantification limit for corticosterone was 1 ng/mL. The validated method was successfully applied to the quantitation of corticosterone in mouse plasma.