顶空固相微萃取/气相色谱-质谱法测定血液中15种挥发性有机物

Determination of 16 Volatile Organic Compounds in Blood by Headspace Solid-phase Microextraction/Gas Chromatography-Mass Spectrometry

该研究使用人工合成模拟血液标准添加溶液,采用部分因子实验、单因素实验、响应曲面法优化萃取条件,建立了顶空固相微萃取/气相色谱-质谱联用(HS-SPME/GC-MS)测定血液中常见挥发性有机物(VOCs)的分析方法。取2.0 mL血液样品,使用65μm聚二甲基硅氧烷/二乙烯基苯(PDMS/DVB)萃取纤维,在萃取温度57℃,萃取时间45 min,搅拌转速1000 r/min,解吸温度280℃、解吸时间5 min、氯化钠含量为0条件下进行萃取,以内标法定量。结果表明,15种VOCs在0.4~4、4~40、40~400 ng/mL质量浓度范围内线性良好(r2>0.99),方法检出限为0.03~0.1 ng/mL,定量下限为0.1~0.3 ng/mL。15种VOCs在2.0、20、200ng/mL加标水平下的平均回收率为71.1%~122%,相对标准偏差(n=6)为7.5%~16%。使用该方法测定新鲜人体血液样品和标准添加样品进行基质效应差异性评价,结果表明模拟血液和真实血液在基质效应方面无显著差异(p>0.05)。该方法线性范围广、检出限和定量下限低、前处理方便、检测门槛低,适用于血液中多种VOCs的检测。

To improve the extraction efficiency of volatile organic compounds(VOCs)in blood,this study employed an artificially synthesized simulated blood standard addition solution as the working solution.Partial factorial experiments,single-factor alternating experiments,and response surface methodology were adopted to optimize the conditions for headspace solid-phase microextraction(HSSPME).An analytical method combining HS-SPME with gas chromatography-mass spectrometry(GC-MS)was established for the determination of common VOCs in blood.The final optimized ex⁃traction method in this study involved the following steps:2.0 mL of blood sample was taken,and 65μm polydimethylsiloxane/divinylbenzene(PDMS/DVB)extraction fiber was used.Extraction was conducted at 57℃for 45 minutes,with stirring speed set at 1000 r/min.Desorption was performed at 280℃for 5 minutes,and the sodium chloride content was set at 0.Internal standard method was used for quantification.The results of methodological validation showed that 15 VOCs exhibited good linearity within the mass concentration ranges of 0.4-4,4-40,and 40-400 ng/mL(r2>0.99).The limits of detection(LODs)of this method ranged from 0.03 to 0.1 ng/mL,and the limits of quantifi⁃cation(LOQs)ranged from 0.1 to 0.3 ng/mL.The average recoveries of 15 VOCs at the mass concen⁃tration levels of 2.0,20,and 200 ng/mL ranged from 71.1%to 122%,and the relative standard de⁃viations(n=6)ranged from 7.5%to 16%.This method was used to determine fresh human blood sam⁃ples and standard addition samples with the same VOCs concentrations to assess matrix effects.The results showed that the p-values of the Shapiro-Wilk test,two-tailed F-test,and two-tailed T-test of the two sets of sample components were all greater than 0.05.The VOCs distributions in both sets of samples conformed to normal distribution,and there were no significant differences in variance and mean values,indicating no significant difference in matrix effects between simulated blood and real blood.The HS-SPME/GC-MS method established in this study possesses wide linear range,low de⁃tection limit and lower limit of quantification,simple pretreatment,and low detection threshold.It is suitable for the detection of multiple VOCs in blood and can provide a basis for subsequent analy⁃sis.