A simple sample preparation approach based on hydrophilic solid-phase extraction coupled with liquid chromatography-tandem mass spectrometry for determination of endogenous cytokinins
A simple sample preparation approach based on hydrophilic solid-phase extraction coupled with liquid chromatography-tandem mass spectrometry for determination of endogenous cytokinins
复制标题
基于亲水性固相萃取结合液相色谱-串联质谱法测定内源细胞分裂素的简单样品制备方法
DOI：
10.1016/j.jchromb.2013.10.024
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发表时间：
2013-12-30
影响因子：
3
通讯作者：
Feng, Yu-Qi
中科院分区：
文献类型：
--
作者：
Cai, Bao-Dong;Zhu, Jiu-Xia;Feng, Yu-Qi
细胞分裂素（CKs）是一类重要的植物激素家族，在植物地上部和根系发育中起着重要的调控作用。然而，植物样品中CK的定量常常受到复杂的植物基质的影响。本研究建立了一种简单、快速、高效的亲水作用色谱-固相萃取（HILIC-SPE）分离纯化肌酸激酶的方法。用乙腈（ACN）提取CK，然后进行HILIC-SPE（硅胶作为吸附剂）纯化。植物样品的提取液可直接用于HILIC-SPE，无需溶剂蒸发步骤，简化了分析过程。此外，HILIC色谱保留机制，疏水性共提取杂质被有效地去除。随后，通过RPLC分离CK，正交于HILIC预处理过程，并通过串联质谱法检测。该方法具有较高的特异性和回收率（> 77.0%）。在0.002 - 100 ng/mL范围内，所有8种肌酸激酶均具有良好的线性关系，相关系数（r）大于0.9927。CK的检测限（LOD，信噪比= 5）在1.0和12.4 pg mL（-1）之间。通过日内和日间测定，对该方法的重现性进行了评价，结果表明，相对标准偏差（RSD）小于10.5%。采用这种方法，我们成功地定量了20 mg水稻叶片中的6种CK，该方法也成功地应用于甘蓝型油菜（花和叶）。(C)2013爱思唯尔有限公司版权所有。
Cytokinins (CKs), a vital family of phytohormones, play important roles in the regulation of shoot and root development. However, the quantification of CKs in plant samples is frequently affected by the complex plant matrix. In the current study, we developed a simple, rapid and efficient hydrophilic interaction chromatography-solid phase extraction (HILIC-SPE) method for CKs purification. CKs were extracted by acetonitrile (ACN) followed by HILIC-SPE (silica as sorbents) purification. The extraction solution of plant samples could be directly applied to HILIC-SPE without solvent evaporation step, which simplified the analysis process. Moreover, with HILIC chromatographic retention mechanism, the hydrophobic co-extracted impurities were efficiently removed. Subsequently, CKs were separated by RPLC, orthogonal to the HILIC pretreatment process, and detected by tandem mass spectrometry. The method exhibits high specificity and recovery yield (>77.0%). Good linearities were obtained for all eight CKs ranging from 0.002 to 100 ng mL(-1) with correlation coefficients (r) higher than 0.9927. The limits of detection (LODs, signal/noise = 5) for the CKs were between 1.0 and 12.4 pg mL(-1). Reproducibility of the method was evaluated by intra-day and inter-day measurements and the results showed that relative standard deviations (RSDs) were less than 10.5%. Employing this method, we successfully quantified six CKs in 20 mg Oryza sativa leaves and the method was also successfully applied to Brassica napus (flower and leaves). (C) 2013 Elsevier B.V. All rights reserved.