Z-Project: Analytical procedures for the determination of the thyroid hormone status in biological samples collected in the framework of the priority programme SPP 1629

Z-Project：确定优先计划 SPP 1629 框架内收集的生物样本中甲状腺激素状态的分析程序

• 批准号: 221224269
• 负责人: Professor Dr. Josef Köhrle
• 金额: --
• 依托单位: Institut für Experimentelle Endokrinologie (CVK)
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2012
• 资助国家: 德国
• 起止时间: 2011-12-31 至 2018-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/221224269?language=en
• 关键词: Project; Analytical; procedures; determination; thyroid

Thyroid hormones (TH) and their metabolites (THM) such as thyronamines (TAM), thyroacetic acids (TA) and several conjugated forms circulate in the blood in free and protein-bound form. During the first SPP funding period two immunoassays (IA) for 3,5-T2 and 3-T1AM in human serum have been optimized, validated and applied for data acquisition in cooperation with SPP 1629 partners. A liquid-liquid extraction (LLE)-based liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for detection of THM profiles in cell culture supernatants has been fully validated and further advanced for cell and tissue extracts using a modified pre-analytical approach involving solid phase extraction (SPE). Optimization and validation of LC-MS/MS methods including pre-analytical sample workup for human and mouse serum is still ongoing. Several experimental limitations in recovery and process efficiency caused by various matrix interferences lead to ion suppression in MS. Radioimmunoassays (RIA) for analysis of rT3 in human serum as well as T4 and T3 in mouse serum have been successfully applied for data generation together with SPP partners. During the second funding period the following aims will be pursued: (1) application of IA to define age and sex-specific reference range(s) for 3T1AM in human serum of children and adults, (2 characterization of development-, gender-, age-, and disease-specific THM profiles in human serum for (personalized) diagnostics and monitoring of therapeutic interventions, (3) completion of validation of THM LC-MS/MS analytics for human and mouse serum once quality control parameters requested by authorities have been accomplished, (4) optimization of pre-analytic TAM extraction from serum-containing matrices. The Z-project as a central platform will provide validated IA and MS-based methods to SPP partners for analytical tasks and quantitative analysis of THM profiles in clinical and experimental samples.

甲状腺激素（TH）及其代谢产物（THM），如甲状腺原氨酸（TAM）、甲状腺乙酸（TA）和几种结合形式，以游离和蛋白结合形式在血液中循环。在SPP的第一个资助期内，与SPP 1629合作伙伴合作，对人血清中3，5-T2和3-T1 AM的两种免疫测定（IA）进行了优化、验证并应用于数据采集。用于检测细胞培养上清液中THM特征的基于液-液萃取（LLE）的液相色谱-串联质谱（LC-MS/MS）方法已得到充分验证，并使用涉及固相萃取（SPE）的改良分析前方法进一步改进了细胞和组织提取物。LC-MS/MS方法的优化和验证（包括人和小鼠血清的分析前样品处理）仍在进行中。几个实验限制回收率和工艺效率所造成的各种基质干扰导致离子抑制MS。放射免疫测定（RIA）分析rT 3在人血清中，以及T4和T3在小鼠血清中已成功地应用于数据生成与SPP合作伙伴。在第二个供资期内，将实现以下目标：（1）应用IA确定儿童和成人血清中3 T1 AM的年龄和性别特异性参考范围，（2）人血清中发育、性别、年龄和疾病特异性THM谱的表征，用于（个性化）诊断和治疗干预的监测，（3）一旦完成当局要求的质量控制参数，就完成了对人和小鼠血清的THM LC-MS/MS分析的验证，（4）优化从含血清基质中提取分析前的TAM。作为一个中心平台，Z项目将为SPP合作伙伴提供经过验证的基于IA和MS的方法，用于临床和实验样品中THM谱的分析任务和定量分析。