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本文建立了白桦(Betula platyphylla)悬浮细胞中S-亚硝基硫醇含量分析的比色法及荧光检测法。其中,比色法采用偶氮化反应;荧光检测法以2,3-二氨基萘(DAN)为探针。比色法结果表明,供试样品在10~35 min内显色稳定性良好,重复性高[S-亚硝基硫醇浓度=(7.54±0.11)μmol·L~(-1),n=6],加样回收率为(101.91±2.75)%(n=4);荧光法结果表明,DAN染色时间和染色浓度均影响白桦悬浮细胞的荧光强度,其中染色时间为2~4 h和染色浓度在400μmol·L~(-1)时细胞荧光强度值最大。由上述结果可知,比色法测定白桦悬浮细胞中S-亚硝基硫醇含量重复性好,准确性高,而荧光检测法直观性强,建议将两方法结合使用分析白桦等植物细胞中S-亚硝基硫醇含量。
In this paper, a colorimetric and fluorescence detection method for the determination of S-nitrosothiol in the suspension cells of Betula platyphylla was established. Among them, the colorimetric method using azo reaction; fluorescence detection method to 2,3-diaminonaphthalene (DAN) as a probe. The results of colorimetry showed that the test samples showed good color stability and reproducibility within 10-35 min [concentration of S-nitrosothiol = (7.54 ± 0.11) μmol·L -1, n = 6] and the recovery rate was (101.91 ± 2.75)% (n = 4). Fluorescence staining showed that both the staining time and the concentration of DAN affected the fluorescence intensity of suspension cells. The staining time was 2 ~ 4 h and the staining time When the concentration was 400μmol·L -1, the fluorescence intensity of cells was the highest. From the above results, the colorimetric determination of S-nitrosothiol content in white birch suspension cells with good reproducibility, high accuracy, and fluorescence detection method is intuitive, it is recommended that the two methods combined with the analysis of plant cells such as birch S - Nitrosothiol content.