National Center for Clinical Laboratories, Beijing Engineering Research Center of Laboratory Medicine, Beijing Hospital, National Center of Gerontology, Institute of Geriatric Medicine, Chinese Academy of Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, P.R. China.
National Center for Clinical Laboratories, Beijing Engineering Research Center of Laboratory Medicine, Beijing Hospital, National Center of Gerontology, Institute of Geriatric Medicine, Chinese Academy of Medical Sciences, Beijing, P. R. China.
Clin Chem Lab Med. 2020 Nov 25;59(3):523-532. doi: 10.1515/cclm-2020-0410. Print 2021 Feb 23.
Accurate measurements of serum 17-hydroxyprogesterone (17OHP) are essential for diagnosis and treatment monitoring for congenital adrenal hyperplasia patients. The performance of serum 17OHP routine methods remains highly variable that calls for a candidate reference measurement procedure (cRMP) to improve the standardization of serum 17OHP measurements.
Serum samples spiked with internal standards were extracted with a combination of solid-phase extraction and liquid-liquid extraction. The 17OHP was quantified by the isotope dilution coupled with liquid chromatography/tandem mass spectrometry (ID-LC/MS/MS) with electrospray ionization in positive ion mode. Nine structural analogs of 17OHP were evaluated for interferences. The precision and analytical recovery were assessed. Twenty native and 40 spiked serum for performance evaluation were measured by the cRMP and two clinical LC/MS routine methods.
No apparent interferences were found with the 17OHP measurement. The within-run, between-run, and total precision for our method were 0.4-0.8%, 0.6-2.0%, and 1.0-2.1% for four pooled serum (2.46-102.72 nmol/L), respectively. The recoveries of added 17OHP were 100.0-100.2%. For the performance of two LC/MS routine methods, they showed relative deviation ranges of -22.1 to 1.1% and -6.7 to 12.8%, respectively.
We developed and validated a reliable serum 17OHP method using ID-LC/MS/MS. The desirable accuracy and precision of this method enable it to serve as a promising cRMP to improve the standardization for serum 17OHP routine measurements.
准确测量血清 17-羟孕酮（17OHP）对于先天性肾上腺皮质增生症患者的诊断和治疗监测至关重要。目前血清 17OHP 的常规方法的性能仍然存在很大差异，因此需要候选参考测量程序（cRMP）来提高血清 17OHP 测量的标准化。
用固相萃取和液液萃取相结合的方法提取添加内标物的血清样品。采用电喷雾正离子模式下的同位素稀释结合液相色谱/串联质谱（ID-LC/MS/MS）定量测定 17OHP。评估了 9 种 17OHP 结构类似物的干扰情况。评估了精密度和分析回收率。采用 cRMP 和两种临床 LC/MS 常规方法测量了 20 份天然血清和 40 份加标血清进行性能评估。
未发现 17OHP 测量有明显干扰。该方法的批内、批间和总精密度分别为四个混合血清（2.46-102.72 nmol/L）的 0.4-0.8%、0.6-2.0%和 1.0-2.1%。添加的 17OHP 的回收率为 100.0-100.2%。对于两种 LC/MS 常规方法的性能，它们的相对偏差范围分别为-22.1%至 1.1%和-6.7%至 12.8%。
我们开发并验证了一种使用 ID-LC/MS/MS 的可靠血清 17OHP 方法。该方法具有良好的准确性和精密度，有望成为提高血清 17OHP 常规测量标准化的有前途的 cRMP。
