Zhang Qiaoxuan, Zhan Min, Wu Huihui, Feng Pinning, Jin Xing, Wan Zemin, Yan Jun, Zhang Pengwei, Ke Peifeng, Zhuang Junhua, Zhou Jiuyao, Han Liqiao, Huang Xianzhang
Department of Laboratory Medicine, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine (Guangdong Provincial Hospital of Chinese Medicine), Guangzhou, China.
Anhui Prevention and Treatment Center for Occupational Disease, Anhui No. 2 Provincial People's Hospital, Hefei, China.
Front Chem. 2022 Aug 11;10:961660. doi: 10.3389/fchem.2022.961660. eCollection 2022.
Accurate investigation of adrenal hormone levels plays a vital role in pediatric endocrinology for the detection of steroid-related disorders. This study aims to develop a straightforward, sensitive UHPLC-MS/MS method to quantify 17 endogenous adrenal corticosteroid hormones in human plasma. These hormones are the main ingredients in the synthetic and metabolic pathways of adrenal corticosteroid hormones. Chromatographic separation was achieved on a C18 column before electrospray ionization triple-quadrupole mass spectrometry in multiple reaction monitoring mode with a run time of 7 min. The samples were extracted by liquid-liquid extraction and required no derivatization. Analytical performance was evaluated, including linearity, analytical sensitivity, accuracy, precision, and specificity. Plasma specimens from 32 congenital adrenal hyperplasia (CAH) patients and 30 healthy volunteers were analyzed to further reveal the diagnostic value of multiple steroid hormones in the synthetic and metabolic pathways of adrenal corticosteroid in CAH diagnosis. All hormones were effectively extracted and separated using our method. The method was essentially free from potential interference of isomers or structural analogues. The imprecisions were <10%. The lower limits of quantification varied from 0.05 to 15.0 ng/ml. Good linearity coefficients ( > 0.998) were also obtained for most hormones in the required concentration range, except for 21-deoxycortisol ( = 0.9967) and androstenediol ( = 0.9952). The recoveries for the steroid hormones ranged from 91.7 to 109.8%. We developed the UHPLC-MS/MS method for the simultaneous measurement of steroid hormones. The results showed that measurement of steroid hormones simultaneously could improve the diagnostic efficiency of CAH.
准确检测肾上腺激素水平在儿科内分泌学中对于类固醇相关疾病的诊断起着至关重要的作用。本研究旨在开发一种简单、灵敏的超高效液相色谱-串联质谱（UHPLC-MS/MS）方法，用于定量测定人血浆中的17种内源性肾上腺皮质激素。这些激素是肾上腺皮质激素合成和代谢途径中的主要成分。在电喷雾电离三重四极杆质谱仪上采用多反应监测模式，在C18柱上进行色谱分离，运行时间为7分钟。样品通过液-液萃取法提取，无需衍生化。对分析性能进行了评估，包括线性、分析灵敏度、准确度、精密度和特异性。分析了32例先天性肾上腺皮质增生症（CAH）患者和30名健康志愿者的血浆标本，以进一步揭示多种类固醇激素在CAH诊断中肾上腺皮质激素合成和代谢途径中的诊断价值。使用我们的方法，所有激素均能有效提取和分离。该方法基本不受异构体或结构类似物的潜在干扰。不精密度<10%。定量下限在0.05至15.0 ng/ml之间。除21-脱氧皮质醇（r = 0.9967）和雄烯二醇（r = 0.9952）外，大多数激素在所需浓度范围内也获得了良好的线性系数（r > 0.998）。类固醇激素的回收率在91.7%至109.8%之间。我们开发了用于同时测定类固醇激素的UHPLC-MS/MS方法。结果表明，同时测定类固醇激素可提高CAH的诊断效率。
