Quantification of succinic acid levels, linked to succinate dehydrogenase (SDH) dysfunctions, by an automated and fully validated liquid chromatography tandem mass spectrometry method suitable for multi-matrix applications
Quantification of succinic acid levels, linked to succinate dehydrogenase (SDH) dysfunctions, by an automated and fully validated liquid chromatography tandem mass spectrometry method suitable for multi-matrix applications
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DOI：
10.1016/j.jchromb.2021.123085
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发表时间：
2022-01-15
影响因子：
3
通讯作者：
Broutin, Sophie
中科院分区：
文献类型：
--
作者：
Lamy, Constance;Mansard, Clemence;Broutin, Sophie
癌症的特征包括代谢与细胞能量失调。琥珀酸脱氢酶（SDH）代谢酶活性异常导致琥珀酸异常积聚已在实体瘤中被描述，但也在炎症和缺血再灌注损伤中被描述。琥珀酸是SDH相关病理诊断、治疗反应评估和疾病随访的潜在生物标志物。我们开发了一种液相色谱串联质谱（LC-MS/MS）方法，可以快速，准确和精确地定量临床（血清，尿液）和临床前（细胞颗粒，上清液）样品中的琥珀酸水平。采用Phenomenex STRATA (TM) XL-A (200 mg - 3 mL) 33 μ m萃取筒进行固相萃取（SPE）前，将C-13(4)琥珀酸二钠盐作为内标加入样品中。该方法由TECAN公司的Freedom EVO (R)平台自动化，琥珀酸在C18色谱柱上与带有电喷雾电离（ESI）源的Xevo (R) TQ-S微型Waters质谱仪结合分离。这种生物医学分析允许标准曲线在1.0-135.5 μ M范围内呈线性，r(2)值为> 0.999，矩阵效应低(
The hallmarks of cancer include metabolism with deregulating cellular energetics. Dysfunctions in succinate dehydrogenase (SDH) metabolic enzyme activity, leading to an abnormal accumulation of succinic acid has been described in solid tumors but also in inflammation and ischemia reperfusion injury. Succinic acid is a potential biomarker of SDH related pathologies for diagnostic, evaluation of treatment response and follow-up of the disease. We developed a liquid chromatography tandem mass spectrometry (LC-MS/MS) method allowing a rapid, accurate and precise quantification of succinic acid levels in clinical (serum, urine) and preclinical (cellular pellets, supernatants) samples. C-13(4) succinic acid disodium salt was used as internal standard and added to samples before a solid phase extraction (SPE) on Phenomenex STRATA (TM) XL-A (200 mg - 3 mL) 33 mu m cartridges. This method is automated by a Freedom EVO (R) platform from TECAN and succinic acid is separated on a C18 column combined to a Xevo (R) TQ-S micro Waters mass spectrometer with electrospray ionization (ESI) source. This biomedical analysis allows standard curves to be linear over the range 1.0-135.5 mu M with r(2) values > 0.999 and low matrix effects (