氨基酸靶标分析用于急性辐射损伤早期伤情分类的初步研究

发布时间：2018-11-09 21:05

【摘要】：核事故后，，对大量辐射暴露人群进行迅速、准确地伤情分类无论对于有限的医疗资源配置还是对大量伤员的分类、处理和早期治疗都显得至关重要。本文首先建立了高效液相色谱/电喷雾串联质谱用于大鼠体液中氨基酸分析的技术平台，然后利用该技术平台从剂量效应和时间效应两方面，考察了急性电离辐射对大鼠体液中氨基酸代谢的影响，并结合非线性偏最小二乘法（PLS）进行了伤情分类的研究。 1．建立了一个基于高效液相色谱/电喷雾串联质谱（HPLC-ESI-MS/MS）与固相萃取（SPE）相结合的用于分析大鼠体液中氨基酸的方法。以硅胶基质的强阳离子交换固相萃取柱（SCX-SPE）提取体液中的氨基酸和降低基质效应，对SPE处理过程中的样品pH值进行了优化。样品pH值为2.8时，SCX-SPE提取氨基酸的回收率和重现性比较令人满意。此外，对分析方法特性进行了考察，包括精密度、准确度及标准曲线等。 2.将建立的液质联用的大鼠血浆中氨基酸分析方法，从剂量效应和时间效应两方面考察了电离辐射对大鼠血浆中氨基酸代谢的影响，结合非线性PLS回归模型进行了伤情分类研究。结果表明，辐射后5h血浆氨基酸对于无辐射对照组的分类正确率为92.3%；辐射后24h，不同程度的伤情分类正确率均在83%以上，其中中度、严重伤情的分类正确率分别达到了91.7%和92.3%；辐射后72h的伤情分类效果最佳，不同程度的伤情分类正确率均达到90%以上，其中中度和极重度的分类正确率达到100%。 3.将建立的液质联用的大鼠尿液中氨基酸分析方法，从剂量效应和时间效应两方面考察了电离辐射对大鼠尿液中氨基酸代谢的影响，结合非线性PLS回归模型进行了伤情分类研究。结果表明，辐射后48h和72h的分类效果优于前面时相点，不同程度辐射损伤的伤情分类正确率在70%以上，但整体分类效果不及血浆样品。
[Abstract]:After the nuclear accident, it is very important to classify a large number of people exposed to radiation quickly and accurately, not only for the limited medical resources allocation, but also for the classification, treatment and early treatment of a large number of injured persons. In this paper, a high performance liquid chromatography (HPLC) / electrospray tandem mass spectrometry (HPLC-MS) platform was established for the analysis of amino acids in rat body fluid. The effects of acute ionizing radiation on amino acid metabolism in the body fluid of rats were investigated, and the classification of injury was studied with nonlinear partial least square method (PLS). 1. A method based on high performance liquid chromatography (HPLC) / electrospray tandem mass spectrometry (HPLC-ESI-MS/MS) combined with solid phase extraction (SPE) was developed for the analysis of amino acids in rat body fluid. The strong cation exchange solid phase extraction (SCX-SPE) column of silica gel matrix was used to extract amino acids from body fluid and to reduce the matrix effect. The pH value of the sample was optimized during SPE treatment. The recovery and reproducibility of amino acids extracted by SCX-SPE were satisfactory when the pH value of the sample was 2.8. In addition, the characteristics of the analytical method, including precision, accuracy and standard curve, are investigated. 2. The effects of ionizing radiation on the metabolism of amino acids in plasma of rats were investigated in terms of dose effect and time effect. The injury classification was studied with nonlinear PLS regression model. The results showed that the classification accuracy of plasma amino acids in the control group without radiation was 92.3% at 5 h after radiation. After 24 hours of radiation, the correct rate of classification of different degrees of injury was above 83%, among which, the correct rate of classification of severe injury was 91.7% and 92.3%, respectively. The classification effect of injury was the best at 72 hours after radiation. The correct rate of injury classification in different degrees was above 90%, and the classification accuracy of degree and severity reached 100%. 3. The effects of ionizing radiation on the metabolism of amino acids in urine of rats were investigated in terms of dose effect and time effect. The injuries were classified with nonlinear PLS regression model. The results showed that the classification effect of 48 h and 72 h after radiation was better than that of the former phase point. The classification accuracy of different degree of radiation injury was more than 70%, but the overall classification effect was not as good as plasma sample.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R818

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