Wang Zhenxin, Wang Hao, Peng Yingfei, Chen Fangjun, Zhao Lin, Li Xiaomu, Qin Jiaqian, Li Qianqian, Wang Beili, Pan Baishen, Guo Wei
Department of Laboratory Medicine, Zhongshan Hospital, Fudan University, Shanghai, P.R. China.
Institute of Biomedical Science, Fudan University, Shanghai, P.R. China.
Clin Chem Lab Med. 2020 Feb 21;58(9):1477-1487. doi: 10.1515/cclm-2019-0869. Print 2020 Aug 27.
Liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based assays are employed in more and more clinical laboratories to quantify steroids. The steroid quantification by LC-MS/MS shows great value in screening or diagnosing endocrine disorders; however, the number of functional steroids included in the LC-MS/MS methods is still limited.
Here, we describe the performance and validation of a 20-steroid plasma panel by LC-MS/MS. The panel included progestogens (including mineralocorticoids and glucocorticoids), androgens and estrogens biosynthesized in steroid metabolic pathways. The LC-MS/MS method was validated according to guidance documents, and subsequently employed to profile steroid changes in endocrine disorders.
Using LC-MS/MS, 20 steroids were separated and quantified in 8 min. Coefficients of variation (CVs) of the 20 analytes at the lower limit of quantification (LLoQ) were all less than 15% (ranging from 1.84% to 14.96%). The linearity of the assay was demonstrated by all the R2 values greater than 0.995. Individual plasma steroids changed significantly in patients with subclinical Cushing's syndrome (SCS) and polycystic ovary syndrome (PCOS) - 17-hydroxypregnenolone (17-OH-PR), testosterone (T) and dihydrotestosterone (DHT) were significantly decreased in SCS patients, while in PCOS patients, pregnenolone, corticosterone (CORT), androstenedione (A4) and T were significantly increased and DHT was decreased.
The LC-MS/MS method we developed for the quantification of 20 plasma steroids is clinical practicable. The steroid profiling data using this assay indicate its screening value for endocrine disorders. To further explore the value of the assay, more investigations are however needed.
基于液相色谱 - 串联质谱（LC-MS/MS）的检测方法在越来越多的临床实验室中用于类固醇的定量分析。通过LC-MS/MS进行类固醇定量分析在筛查或诊断内分泌疾病方面具有重要价值；然而，LC-MS/MS方法中所包含的功能性类固醇数量仍然有限。
在此，我们描述了一种通过LC-MS/MS检测20种类固醇的血浆检测方法的性能及验证情况。该检测组合包括在类固醇代谢途径中生物合成的孕激素（包括盐皮质激素和糖皮质激素）、雄激素和雌激素。LC-MS/MS方法根据指导文件进行了验证，随后用于分析内分泌疾病中的类固醇变化情况。
使用LC-MS/MS，在8分钟内分离并定量了20种类固醇。20种分析物在定量下限（LLoQ）处的变异系数（CVs）均小于15%（范围为1.84%至14.96%）。所有R2值均大于0.995，证明了该检测方法的线性。亚临床库欣综合征（SCS）和多囊卵巢综合征（PCOS）患者的个体血浆类固醇发生了显著变化——SCS患者中17-羟孕烯醇酮（17-OH-PR）、睾酮（T）和双氢睾酮（DHT）显著降低，而在PCOS患者中，孕烯醇酮、皮质酮（CORT）、雄烯二酮（A4）和T显著升高，DHT降低。
我们开发的用于定量20种血浆类固醇的LC-MS/MS方法在临床上是可行的。使用该检测方法获得的类固醇分析数据表明其在筛查内分泌疾病方面的价值。然而，为了进一步探索该检测方法的价值，还需要更多的研究。
