摘要
基于IKnife-快速蒸发离子化质谱技术对低脂鱼类南极犬牙鱼的肌肉组织进行脂质组学轮廓检测,并通过响应面分析优化系统参数。结果表明,最优参数为输出功率25 W、切割速率0.50 mm/s、切割长度1.0 cm。在最佳实验条件下共检出脂肪酸离子17种,其中信号响应强度最大为m/z 327.23,相对含量达到了17.81%,经鉴定为脂肪酸(fatty acid,FA),结构为FA 22∶6,其次分别为m/z 255.23(FA 16∶0,9.81%)和m/z 281.25(FA 18∶1,9.09%);检出磷脂离子10种,质量范围为m/z 736.49~909.55,信号最强离子峰m/z 885.55经鉴定为磷脂酰肌醇(phosphatidylinositol,PI)离子[PI 38∶4-H]-,相对丰度达到了19.30%,其次为磷脂酰乙醇胺(phosphatidylethanolamine,PE)离子m/z 790.54([PE 40∶6-H]-,15.24%)。方法验证结果显示,目标离子的信噪比为35.4~95.3,偏差为4.17×10~(-6)~9.78×10~(-6),日内精密度相对标准偏差为3.7%~5.6%,日间精密度相对标准偏差为5.9%~7.3%。本实验方法灵敏度和分辨率高、检测速度快、结果稳定,为脂质组学研究和鱼类生物信息检测提供了新的技术手段。
Rapid evaporative ionization mass spectrometry(REIMS) was employed for lipidomic profiling of Patagonian toothfish, a kind of low-fat fish. The instrumental parameters were optimized using response surface methodology with a Box-Behnken design as follows: power output 25 W, cutting speed was 0.50 mm/s, and cutting length 1.0 cm. Under these optimized conditions, a total of 17 fatty acid molecular species were detected, among which the signal intensity of m/z 327.23(FA 22:6, 17.81%) was the highest, followed by m/z 255.23(FA 16:0, 9.81%), and m/z 281.25(FA 18:1, 9.09%).A total of 10 phospholipid molecular species were tentatively identified in the m/z range from 736.49 to 909.55. The peak at m/z 885.55 was the most intensive with a relative content of 19.30%, which was identified as phosphatidylinositol(PI)species [PI 38:4–H]–, followed by phosphatidylethanolamine(PE) species at m/z 790.54([PE 40:6–H]–, 15.24%). The method was validated and the results showed that the signal-to-noise ratios of the target ions were in the range of 35.4–95.3,with error values of 4.17 × 10~(-6)–9.78 × 10~(-6). The relative standard deviations(RSDs) of intra-day and inter-day precision were 3.7%–5.6% and 5.9%–7.3%, respectively. The results indicate that this method is sensitive and stable with high resolution. It provides a totally new technology for lipidomic and bioinformatics analysis offish.
引文
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