LC-MS同时测定大型海藻中9个植物激素

目的: 利用高效液相色谱-三重四极杆质谱(HPLC-QqQ MS)联用分析系统建立几种大型海藻中植物激素的分析方法。 方法: 冷冻干燥后的藻样用甲醇-水-甲酸(15∶4∶1,v/v/v)提取。9个中、酸性植物激素分别在Hypersil Gold C₁₈色谱柱上实现分离,流动相为水(A)-甲醇(B),梯度洗脱,流速0.3 mL·min^-1,柱温30 ℃。在电喷雾电离源正离子模式下,分别取m/z 130.0,204.1,136.1,220.0作为吲哚乙酸、异戊烯腺苷、异戊烯腺嘌呤和反式玉米素核苷的定量目标离子;在负离子模式下分别取153.3,91.3,93.2,143.2,59.5作为脱落酸、苯乙酸、水杨酸、赤霉素和茉莉酸的定量目标离子进行检测。 结果: 植物激素浓度在5~500 μg·L^-1范围内线性良好,检测限为0.0041~0.8622 μg·L^-1,回收率在61.33%~90.39%之间,RSD＜11%(n=3)。 结论: 本法操作简单,准确可靠,适用于藻类植物激素的含量检测。

Objective: To develop a method for the determination of phytohormones in several macroalgaes. Methods: The tissues were extracted with methanol-water-formic acid(15∶4∶1,v/v/v).Nine kinds of phytohormones were separated with high efficiency in the selective reaction monitoring(SRM) mode on Hypersil Gold C₁₈ column,with the mobile phase consisting of water(A) and methanol(B) in a programmed gradient elution.The flow rate was 0.3 mL·min^-1 and the column temperature was 30 ℃.The mass spectrometry was performed using precursor ions at m/z 130.0,204.1,136.1 and 220.0 as quantitative ions in positive mode for indole-3-acetic acid,N⁶-(2-isopentyl) adenosine,N⁶-(2-isopentenyl) adenosine and trans-zeatin riboside,respectively.Meanwhile,in negative mode,m/z 153.3,91.3,93.2,143.2 and 59.5 were used as quantitative ions for abscisic acid,phenylacetic acid,salicylic acid,gibberellin A₃ and jasmonic acid. Results: An excellent linear relationship was achieved in the range of 5-500 μg·L^-1,detection limit of phytohormones was from 0.0041 μg·L^-1 to 0.8622 μg·L^-1,the recoveries varied between 61.33%-90.39% with RSD＜11%(n=3). Conclusion: This method of the operation is accurate,reliable and can be used for the phytohormones content determination in macroalgaes.