Analysis of the Structural-related Impurities Involved in 3,3',5-Triiodo-L-thyronine (T3) Purity Reference Material
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摘要: 研制三碘代-L-甲状腺原氨酸(T3)纯度标准物质,可为临床测量结果提供溯源源头,为甲状腺疾病的精准诊断提供可靠依据。在质量平衡法定值过程中,对候选物结构相关杂质进行化学结构鉴定与定量,是降低主成分定值结果不确定度的有效方式。以核磁共振、高分辨质谱为化学分析工具,首先确认了T3标准物质主成分的结构;随后基于高效液相色谱、高分辨质谱、离子淌度等分析技术开发建立了结构相关杂质的分析方法,对T3纯度标准物质中的有机杂质进行分离、分析、鉴定。结果表明,NIM-RM3239标准物质中相对含量>0.1%的结构类似杂质共3种,分别为二碘代-L-甲状腺原氨酸、氯代三碘代-L-甲状腺原氨酸、L-甲状腺素,对映异构体拆分结果表明NIM-RM3239中不含主成分的对映异构体。研究结果可作为质量平衡法定值依据,用于研制纯度型T3国家一级有证标准物质。Abstract: Research and development of 3,3',5-Triiodo-L-thyronine (T3) purity reference material is a way to establish the primary material for clinical measurement value traceability, which can provide more accurate and comparable evidence in thyroid disease diagnosis. In the mass balance quantitative approach, determining and analyzing those structural-related impurities is essential. Such investigation could also be an effective way to reduce the uncertainty of the standard value. First, nuclear magnetic resonance and high-resolution mass spectrometry analytical tools have been employed to validate the chemical structure of the reference material. Then, the structural-related impurities are separated and qualitatively analyzed by using reliable analytical technologies, including high-performance liquid chromatography, high-resolution mass spectrometry, and ion mobility spectrometry. The results indicated that there are three kinds of structural-related impurities involved in the T3 purity reference material (NIM-RM3239), which are Diiodo-L-thyronine, Chloro-diiodo-L-thyronine, and L-Thyroxine. The relative content of each impurity is greater than 0.1% in UV 220nm. Furthermore, the chiral isomer separation result indicated that no dextroisomer (D-T3) existed in NIM-RM3239. These analytical results can be utilized as the foundations in the mass balance approach, applied in developing T3 primary certified reference material.
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图 6 L-/D-T3的离子淌度质谱图(A,B)波速300 m/s、波高40v下淌度谱图;(C,D)波速900 m/s、波高40v下淌度谱图;(E,F)波速300 m/s、波高40v下淌度谱图
Figure 6. Ion mobility spectrometry of L-/D-T3 solution under (A,B)wave velocity 300 m/s, wave height 40v; (C,D)wave velocity 900 m/s, wave height 40v, and (E,F)wave velocity 1200 m/s, wave height 40v, respectively
表 1 T3纯度标准物质中主要结构近似杂质分布
Table 1. The main impurities involved in T3 reference material
代号 质荷比 分子式 杂质1 525.89 C15H13I2NO4 杂质2 559.86 C15H11ClI2NO4 杂质3 777.69 C15H11I4NO4 -
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