In the present study an ultra-performance convergence chromatography tandem mass spectrometry (UPC²-MS/MS) analytical method was developed and validated for the determination of 17 C₁₉ and 14 C₂₁ steroids, including C11-oxy C₁₉ and C11-oxy C₂₁ steroids. The limit of detection and limit of quantification ranged from 0.01 to 10 ng/mL and from 0.01 to 20 ng/mL, respectively, and the method shows the recovery, matrix effect and process efficiency of steroids isolated from a serum matrix to be within acceptable limits. Good accuracy, repeatability and reproducibility were also shown and the method provided excellent sensitivity and selectivity as stereoisomers and regioisomers were also resolved and quantified accurately.

Clinical conditions such as congenital adrenal hyperplasia, polycystic ovary syndrome in females and disorders of sex development in neonates and in children, amongst others, are characterized by abnormal steroid levels. Steroid profiling is essential to accurately diagnose steroid levels in the above settings as well as in androgen excess or deficiency in adrenal-linked endocrine diseases. Our method, separating C₁₉ and C₂₁ steroids in a single chromatographic step, offers a reduced sample turnover rate in the clinical setting, while providing comprehensive steroid profiles of in vivo steroids in the nmol/L range. This is, to our knowledge, the first method reported to simultaneously separate C₁₉ and C₂₁ steroids, together with their C11-hydroxy and C11-keto metabolites –one which may hold promise in the identification of new steroid markers in steroid-linked endocrine diseases, in addition to profiling steroid metabolism and abnormal enzyme activity in patients.

在本研究中，开发了一种超高效能色谱串联质谱分析法（UPC ² -MS / MS）并已验证用于测定17 C ₁₉和14 C ₂₁类固醇，包括C11-氧基C ₁₉和C11-氧基C ₂₁类固醇。检测限和定量限分别为0.01至10 ng / mL和0.01至20 ng / mL，该方法显示从血清基质中分离的类固醇的回收率，基质作用和加工效率均在可接受范围内限制。还显示了良好的准确性，可重复性和重现性，并且该方法提供了出色的灵敏度和选择性，因为立体异构体和区域异构体也得到了准确解析和定量。

诸如先天性肾上腺增生，女性多囊卵巢综合征以及新生儿和儿童性发育障碍等临床症状的特征是类固醇水平异常。在上述情况下以及在雄激素过多或缺乏的肾上腺相关内分泌疾病中，类固醇概况分析对于准确诊断类固醇水平至关重要。我们的方法在单个色谱步骤中分离C ₁₉和C ₂₁类固醇，在临床环境中降低了样品的转化率，同时提供了nmol / L范围内体内类固醇的全面类固醇概况。据我们所知，这是第一种同时分离C ₁₉和C _21的方法 类固醇以及它们的C11-羟基和C11-酮代谢物–除了对类固醇代谢和异常酶活性进行谱分析外，还可以在类固醇相关内分泌疾病中鉴定新的类固醇标志物。