基于HPLC-QqQ-MS/MS技术的坛紫菜中植物激素分析

利用高效液相色谱-质谱联用技术对不同时期和不同品种坛紫菜中的植物激素进行分析研究。通过优化植物激素的提取方法，最终确定采用甲醇-水-甲酸（15∶4∶1，V/V）作为提取溶剂超声提取坛紫菜中的9 种植物激素，选用10 mmol/L乙酸铵的甲醇-水-乙酸（90∶10∶0.05，V/V）作为复溶溶剂。此外经过流动相、固定相、复溶试剂等方面的条件选择，确定采用Thermo Hypersil Gold C18色谱柱（100 mm×2.1 mm，3 μm）为固定相，以含10 mmol/L乙酸铵溶液-甲醇为流动相，在选择反应监测模式下对吲哚乙酸、异戊烯腺苷、异戊烯腺嘌呤、反式玉米素核苷、玉米素、脱落酸、芸苔素内酯、水杨酸和赤霉素进行含量分析。结果表明，9 种激素标准品线性关系良好，相关系数（R2）均高于0.991，紫菜内主要激素（除赤霉素）回收率均在71%以上；方法的检出限均低于3 μg/L，定量限在0.45～5.0 μg/L之间。利用该方法对坛紫菜进行激素含量分析，比较分析了不同品种坛紫菜的激素分布以及不同时期（一水、二水和三水）坛紫菜中9 种植物激素的变化规律，分析了激素含量与坛紫菜形态学相关性，了解坛紫菜的激素分布情况。

Simultaneous Analysis of Nine Phytohormones in Porphyra haitanensis by High Performance Liquid Chromatography Coupled with Triple Quadrupole Tandem Mass Spectrometry

A quantitative method based on high performance liquid chromatography-tandem mass spectrometry
(HPLC-MS) has been established for simultaneous analysis of nine phytohormones in Porphyra haitanensis. The optimized
sample preparation procedure involved extraction with methonal-H2O-formic acid (15:4:1, V/V) and re-dissolution in
methonal-H2O-acetic acid (90:10:0.05, V/V) containing 10 mmol/L ammonium acetate. The chromatography was performed
using a Thermo Hypersil Gold C18 column (100 mm × 2.1 mm, 3 μm) as the stationary phase, and the mobile phase consisted
of aqueous solution containing 10 mmol/L ammonium acetate-methanol. Nine phytohormones (indole-3-acetic acid, N6-(2-
isopentyl) adenosine, N6-(2-isopentenyl)adenosine, trans-zeatin riboside, zeatin, abscisic acid, salicylic acid, gibberellin A3,
and brassinolide) were quantified using selective reaction mode. As results, good linear relationships between concentrations
and the corresponding peak areas of nine standards were achieved with correlation coefficients (R2) higher than 0.991. The
recoveries of eight standards except gibberellin A3 were higher than 71%. The limits of detection (LODs) of phytohormones
by this method were all lower than 3 μg/L and the limits of quantification (LOQs) were in the range from 0.45 to 5 μg/L.
Using the established method, the concentrations of nine phytohormones in different varieties of Porphyra haitanensis and at
different cultivation periods were successfully investigated and correlated with its morphology.