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目的:对化学发光法测定血浆一氧化氮水平的样品前处理方法进行优化,增强该检测方法的精确度及可操作性。方法:①应用酒精对血浆样品进行预处理,比较与去离子水预处理在检测样本通量、信噪比和检测结果方面的差异;②观察不同加样体积对检测结果的影响;③应用优化后的方法检测大鼠血浆一氧化氮参考值范围。结果:①应用酒精预处理样品使得一次检测的样本数量及信噪比均高于去离子水处理;②当加样体积<30μl时,随加样体积的减小,一氧化氮检测结果偏离真实值的幅度呈逐渐增加趋势,>30μl时则与真实值趋于接近;③禁食大鼠动脉血浆一氧化氮浓度范围为5.82～27.38μmol/L。结论:①用酒精代替去离子水预处理样品在保持检测精确度的同时可以提高检测信噪比,增加一次检测的样本数量;②样品加样体积对检测结果的准确性存在较大影响,加样体积应>30μl;③优化后的化学发光法检测的大鼠血浆一氧化氮浓度范围与其他方法检测结果一致,提示方法具有良好的可靠性和准确性。
OBJECTIVE: To optimize the sample preparation method for the determination of plasma nitric oxide by chemiluminescence and to improve the accuracy and operability of this method. Methods: ① Plasma samples were pretreated with alcohol, and compared with the pretreatment of deionized water in the sample throughput, signal-to-noise ratio and test results differences; ② observed the impact of different sample volume on the test results; ③ application optimization After the method of detection of plasma nitric oxide reference range. Results: ① The amount of sample and the signal-to-noise ratio of a sample pretreated with alcohol were both higher than that of the deionized water; ② When the sample volume was less than 30μl, the nitric oxide test results deviated from the real The amplitude of the value increased gradually, while the value of> 30μL tended to be close to the true value. ③ The concentration of nitric oxide in the plasma of fasted rats was 5.82-27.38μmol / L. Conclusion: (1) Pretreatment of samples with alcohol instead of deionized water can improve the detection signal-to-noise ratio (SNR) and increase the number of samples to be tested at one time while maintaining the accuracy of detection. (2) The sample volume added has a great influence on the accuracy of the detection results. Sample volume should be> 30μl; ③ optimized chemiluminescence detection of plasma concentrations of nitric oxide in rats and other methods consistent with the test results, suggesting that the method has good reliability and accuracy.