Livie Craig, Lee Louisa C Y, Alimussina Malika, Conceicao Carina, Leung Chui Ha, McNeilly Jane, Ahmed Syed Faisal, Syme Charlotte, Smith Karen, Johnston Susan
Department of Clinical Biochemistry, Glasgow Royal Infirmary, Glasgow, UK.
Department of Clinical Biochemistry, Royal Alexandra Hospital, Paisley, UK.
J Chromatogr B Analyt Technol Biomed Life Sci. 2023 Apr 1;1220:123615. doi: 10.1016/j.jchromb.2023.123615. Epub 2023 Jan 31.
The measurement of dehydroepiandrosterone-sulphate (DHEAs) is an important second-line test to aid in the diagnosis of premature adrenarche, peripubertal gynaecomastia in males and in identifying the source of elevated androgens in females. Historically, DHEAs has been measured by immunoassay platforms which are prone to poor sensitivity and more importantly poor specificity. The aim was to develop an LC-MSMS method for the measurement of DHEAs in human plasma and serum, develop an in-house paediatric (<6 year old) reference limit and compare the performance against the Abbott Alinity DHEAs immunoassay method. Following pre-treatment with an internal standard, samples were loaded onto EVOLUTE® EXPRESS ABN plate. Analytes were separated with reverse-phase chromatography using ACQUITY® UPLC® HSS T3 2.1 mm × 50 mm, 1.8 μm column. Mass spectrometry detection was performed using a Waters® Xevo TQ-XS in electrospray negative mode. For the paediatric reference range, samples were collected from an inpatient setting (age ≤ 6 years old) with no evidence of adrenal dysfunction or history of/current steroid use. The method comparison was performed using samples from this cohort aged between 0 and 52 weeks. The assay demonstrated linearity up to 15 µmol/L (r > 0.99) with a functional sensitivity of 0.1 µmol/L. Accuracy results revealed a mean bias of 0.7% (-14% to 15%) when compared against the NEQAS EQA LC-MSMS consensus mean (n = 48). The paediatric reference limit was calculated as ≤ 2.3 µmol/L (95% C.I. 1.4 to 3.8 µmol/L) for ≤ 6 year olds (n = 38). Comparison of neonatal (<52 weeks) DHEAs with the Abbott Alinity revealed that the immunoassay ran at a 166% positive bias (n = 24) which appeared to lessen with increasing age. Described is a robust LC-MSMS method for the measurement of plasma or serum DHEAs validated against internationally recognised protocols. Comparison of paediatric samples of <52 weeks against an immunoassay platform demonstrated that in the immediate new-born period results generated from the LC-MSMS method offer superior specificity than an immunoassay platform.
硫酸脱氢表雄酮（DHEAs）的测定是一项重要的二线检测，有助于诊断性早熟、男性青春期前后乳腺增生以及确定女性雄激素升高的来源。历史上，DHEAs一直通过免疫分析平台进行测定，这些平台灵敏度较差，更重要的是特异性较差。目的是开发一种用于测定人血浆和血清中DHEAs的液相色谱-串联质谱（LC-MSMS）方法，制定内部儿科（<6岁）参考限值，并将其性能与雅培Alinity DHEAs免疫分析方法进行比较。用内标进行预处理后，将样品加载到EVOLUTE® EXPRESS ABN板上。使用ACQUITY® UPLC® HSS T3 2.1mm×50mm、1.8μm色谱柱通过反相色谱法分离分析物。使用Waters® Xevo TQ-XS在电喷雾负离子模式下进行质谱检测。对于儿科参考范围，从无肾上腺功能障碍证据或无类固醇使用史/当前无类固醇使用的住院患者（年龄≤6岁）中采集样本。使用该队列中年龄在0至52周之间的样本进行方法比较。该检测方法在高达15µmol/L时呈线性（r>0.99），功能灵敏度为0.1µmol/L。与NEQAS EQA LC-MSMS共识均值（n = 48）相比，准确度结果显示平均偏差为0.7%（-14%至15%）。≤6岁儿童（n = 38）的儿科参考限值计算为≤2.3µmol/L（95%置信区间1.4至3.8µmol/L）。将新生儿（<52周）DHEAs与雅培Alinity进行比较发现，免疫分析的阳性偏差为166%（n = 24），且该偏差似乎随着年龄增长而减小。本文描述了一种稳健的LC-MSMS方法，用于测定血浆或血清中的DHEAs，并根据国际认可的方案进行了验证。将<52周的儿科样本与免疫分析平台进行比较表明，在新生儿早期，LC-MSMS方法产生的结果比免疫分析平台具有更高的特异性。
