分散固相萃取-液相色谱-质谱法测定水稻叶片中的10种代谢物

Determination of 10 kinds of metabolomics in rice shoot by dispersive solid phase extraction-liquid chromatography-mass spectrometry

中文关键词:  代谢物  分散固相萃取  水稻叶片  液相色谱-质谱法

英文关键词:metabolomics  dispersive solid phase, rice shoot  liquid chromatography-mass spectrometry

中文摘要:

      目的 建立分散固相萃取-液相色谱-质谱法(liquid chromatography-mass spectrometry, LC-MS)测定水稻叶片中10种代谢物的分析方法。方法 水稻叶片样品经甲醇-乙腈-水溶液(2:2:1, V:V:V)提取, 无水硫酸钠盐析, 以乙二胺-N-丙基硅烷(primary secondary amine, PSA)和石墨化炭黑(graphitized carbon black, GCB)作为分散吸附剂进行净化, 最后进入液相色谱-质谱仪进行检测。结果 方法在12 min内完成了10种代谢物的定量分析。在3个添加水平(0.50、10.00、100.00 ng/L)下, 水稻叶片中10种代谢物的平均回收率为75.1%~122.2%, 相对标准偏差(relative standard deviations, RSDs)为3.8%~8.9% (n=6)。10种代谢物的标准工作曲线相关系数(r)均大于0.99879, 检出限(limits of detection, LODs)为0.016~0.039 ng/L。结论 本方法具有相对简便、节省时间、灵敏度较高等优势, 可以用于水稻叶片中代谢产物的定量检测。

英文摘要:

      Objective To establish a method for the determination of 10 kinds of metabolites in rice shoot by dispersive solid phase extraction-liquid chromatography-mass spectrometry (LC-MS). Methods Rice shoot samples were extracted by methanol-acetonitrile-aqueous solution (2:2:1, V:V:V), salted out by anhydrous sodium sulfate, ethylenediamine-N-propyl silane (PSA) and graphitized carbon black (GCB) were used as dispersive adsorbent for purification, and then detected by LC-MS. Results The quantitative analysis of 10 kinds of metabolites was completed within 12 min, the average recoveries of 10 kinds of metabolomics in rice shoot were 75.1%?122.2% at three spiked levels (0.50, 10.00, 100.00 ng/L), and the relative standard deviations (RSDs) were from 3.8% to 8.9% (n=6). The calibration curves of 10 kinds of metabolomics in rice shoot showed correlation coefficients (r) higher than 0.99879, and the limits of detection (LODs) were 0.016?0.039 ng/L. Conclusion The method is relatively simple, time-saving and high sensitivity, and is expected to be used for quantitative determination of metabolites in rice shoot.