【中文摘要】人体血清中葡萄糖、氢化可的松的含量测定是临床检测中极为关键的生化指标之一,为临床诊断和病情观察提供了可靠的依据,对相关疾病的诊断和治疗具有重要意义。目前已有多种方法应用于临床实验室中血清葡萄糖与氢化可的松的测定,但由于其测定结果间缺乏可比性,因此实现测定标准化迫切需要建立正确、精密的参考方法。本文中以D-[~(13)C_6]-葡萄糖作为丈量内标,分别考查了血清葡萄糖正确测定所需的样品前处理、衍生化反应、色谱质谱的分离和丈量条件,建立了同位素稀释质谱技术正确测定人体血清中葡萄糖的简单方法。文中采用本法对三种不同浓度血清中葡萄糖的含量进行定值,丈量结果的相对标准偏差均小于1%;同时采用本法对标准物质SRM965a(level1、level 3)冰冻人血清进行了葡萄糖的方法正确性的验证,与标准物质保证值的相对偏差均在±1%以内;同时,进行了回收率实验,方法的回收率在99～101%之间,证实方法正确度较高。同时本文对葡萄糖的定值结果进行了全面的不确定度评定,总的不确定度在1%以内,符合人体血清葡萄糖测定的参考方法的要求。本文提出的气相色谱—同位素稀释/质谱法测定人体血清中葡萄糖的测定方法,前处理过程比较简单,是较为正确的分析方法,可作为人体血清葡萄糖测定的参考方法。同位素稀释质谱技术同样应用于测定人体血清中氢化可的松。2006年,中国计量科学研究院化学计量与分析科学研究所参加了国际计量局物质量咨询委员会(CCQM)组织血清氢化可的松的国际比对CCQM-P77a,应用液相色谱—同位素稀释/质谱法测定,方法精密、正确,具有良好的特异性和灵敏度,测定结果与其他各国达到等效一致。');
【Abstract】 Glucose and cortisol in serum is steroid hormone that is involved in the regulation of the menstrual cycle.Serum glucose and cortisol measurements are routinely performed with different techniques which are *** and sensitive but susceptible to various sources of interference.Thus there is a strong need to establish a critically evaluated reference measurement.In this thesis,GC-ID/MS method was developed to determine serum glucose.D-[~(13)C_6]-glucose was selected as labeled materials in the research. The conditions of serum sample pretreatment,derivatization and the operation parameters of GC-MS were studied.At the optimized conditions,the RSD of the glucose at three levels for human serum samples by the GC-ID/MS was less than 1%.Two NIST standard samples SRM 965a(Level 1,Level 3)were used as certificated standards to evaluate the established method.The result showed that relative diviationes were less than±1%.The analytical recoveris ranged from 99%to 101%.The total uncertainty of glucose of four human serum samples assays were less than 1%.The results showed that it could be hopefully recommended as reference method for glucose assays.The ***,reliable and high-accuracy GC-ID/MS method for serum glucose has been developed.The method may also be used as the standard and reference method of glucose assay in serum samples.The isotope dilution/Mass spectrum(ID/MS)technique was also developed to determine serum cortisol.In 2006,with the LC-ID/MS method, National Institute of Metrology/National Research Center for CRMs (NIM/NRCCRM)participated in the international comparison CCQM-P77a (determination of serum cortisol)held by the Consultative Committee for Amount of Substance(CCQM).Because that the method was accurate,precise and sensitive and our results agreed well with that reported by other participating national metrology institutes.
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